AccuPower® PreMix PCR master mix is available with or without loading dye. When loading dye is present you can load it directly into your gel following PCR – no glycerol or glucose is required.
 

Features and Benefits

•  Fast: just add primer and template
•  Convenient: no more dNTP or enzyme stock solutions required
•  Flexible: available in tube (0.2 or 0.5 ml) or plate (96 or 384 well) formats
•  Stable: for up to 2 years in the freezer and one month at room temperature
•  Customizable: available with primers already added for even greater convenience
•  Gel loading : available with or without tracking dye for ease of use
   

RNA Amplification

Specifications of AccuPower PreMix series

Bioneer offers a wide range of enzymes for PCR, Reverse Transcription, Reverse Transcriptase PCR (RT-PCR) and DNA Ligation. Our Top DNA Polymerase is the fastest thermostable PCR polymerase on the market. We are the only company to use a novel “enzyme-mediated” HotStart in our enzymes. In addition, our CycleScript Reverse Transcriptase uses a Cyclic-RT reaction that allows for robust cDNA synthesis of the rarest transcripts.

For faster and easier reaction set up, the AccuPower® PreMix Series contains each enzyme and all other PCR contents in an easy to resuspend lyophilized mix.

 

Features and Benefits

• Fast: Highly processive enzymes

• Flexible: for all applications from standard, to high fidelity and long range and more!

• Value: No license fees associated with our novel enzymes saves you money!

Specifications of Bioneer Enzymes

Bioneer’s AccuPower® PCR PreMix is a convenient lyophilized PCR master mix containing Top DNA Polymerase, dNTPs, reaction buffer, tracking dye, and a patented stabilizer. AccuPower® PreMix PCR master mix includes our super-processive “three times faster than Taq” Top DNA Polymerase for the fastest nucleic acid amplification.
While Top DNA Polymerase is ideal for applications where you would normally use Taq, if your needs include HotStart PCR or High Fidelity PCR, please see our AccuPower® HotStart PCR and AccuPower® TLA product pages. AccuPower® PreMix PCR master mix is available with or without tracking dye, depending on your application.
If purchased with tracking dye, reactions can be loaded onto agarose gels without adding loading buffer.

Features and Benefits

• Ease of use: just add template and primers and start your PCR. dNTPs, buffer and enzyme are incorporated.
• Stability: stable at room temperature for a month and up to 2 years in a -20°C freezer
• Gel loading: available with or without a tracking dye for ease of use

5' to 3' exonuclease activity: No
3' to 5' exonuclease activity: No
Terminal transferase activity: Yes
Fragment Size: Up to 10 kb

Application

Standard PCR



Figure 1. Processivity comparison with Lambda DNA template and different PCR reagents.

M: 100 bp Plus DNA Ladder (D-1035)
A: AccuPower® PCR PreMix
B: Taq mastermix from Company S
C: Taq mastermix from Company T
Lane M: 1 kb DNA Ladder (D-1040)
Lane 1: 1 kb PCR product
Lane 2: 2 kb PCR product
Lane 3: 3 kb PCR product
Lane 4: 4 kb PCR product


Figure 2. Comparison of sensitivity test for AccuPower® PreMix PCR master mix and other companies' products using serial diluted human genomic DNA.
Reaction condition: 95°C for 5 minutes, followed by 35 cycles of 20 seconds at 95°C, 20 seconds at 60°C, 30 seconds at 72°C.

A: human Insulin receptor gene
B: DNA cross-link repair 1A gene
I: AccuPower® PCR PreMix
II: Taq DNA Polymerase from Company A
III: Taq DNA Polymerase from Company Q
IV: PCR PreMix from Company I
M: 100 bp DNA ladder (D-1030)
Lane 1: 10 ng
Lane 2: 1 ng
Lane 3: 100 ng
Lane 4: 10 ng

• AccuPower® PCR PreMix

MSDS

• MSDS_AccuPower® PCR PreMix



Brochure

• AccuPower® PreMix series - 2010 Brochure

Quality Assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

The AccuPrep® DNA/ RNA Extraction Kit uses a single spin column with glass micro fiber filters.
Bioneer provides various products for high-purity and high-yield Genomic DNA, Plasmid DNA, Viral RNA or fragment DNA. TheAccuPrep® Nano-Plus Plasmid DNA Extraction Kit uses Bioneer’s proprietary cell fragment and protein removal technology (KOR: 10-085043, Nucleic acid separation method and nucleic acid separation mixture using particle material) to reduce plasmid DNA prep time from the usual 30 minutes to a revolutionary 10 minutes. The nano-particles contained within the product form a complex with cellular debris, denatured proteins and chromosomal DNA to form a clear lysate that contains only plasmid DNA within just 1 minute (compared to the traditional 10 minutes), shortening the entire process to a mere 10 minutes.

The AccuPrep® DNA Extraction Kit for 96 well vacuum manifold is designed for high-throughput DNA extraction. The 96-well plate format kit is attached to a vacuum manifold (BioVac™ 96 vacuum manifold) to extract and purify DNA from up to 96 samples simultaneously.

The AccuPrep® Quick DNA Kit enables you to extract PCR-grade genomic DNA by incubation in a single tube. The product is based on Bioneer’s proprietary cell fragment and protein removal technology (KOR: 10-085043, Nucleic acid separation method and nucleic acid separation mixture using particle material) to extract genomic DNA from samples such as mouse or rat tail tips without the complex workflow commonly associated with this type of extraction.

Finally, the PrepMate and AccuZol products use traditional organic solvent DNA/RNA extraction methods. These products contain phenol and chaotrotic salts to stop the activity of various nucleases and extract high-quality nucleic acids. These products are especially suitable for large input volumes or a manual workflow.

Bioneer, founded in 1992, is one of the world’s leading suppliers of synthetic oligonucleotides (DNA and RNA).
In-house production of raw materials (phosphoramidites, reagents & solvents) combined with automated oligo synthesis and purification systems, results in superior quality oligonucleotides at a great price.
Since Bioneer manufactures all of the components that go into our oligonucleotides, we manage quality control every step of the way – ensuring that you receive only the highest quality product.
Every Bioneer oligonucleotide is purified free of charge utilising our unique Bio-RP cartridge purification technology. Bio-RP removes many impurities and synthesis failure products that are still present in an oligo after desalting*.
These impurities contribute to the OD measurement of competitor oligonucleotides that are merely desalted – and artificially inflate yield (see below).
Bioneer’s standard Bio-RP cartridge purification is near HPLC quality with only full-length product resulting. 
We are so confident with our oligonucleotide quality that we are the only company to offer a 100% guarantee that they will work for your PCR or qPCR application!
With high throughput oligo synthesis facilities around the world, Bioneer’s daily capacity is unsurpassed.
Bioneer is unrivalled in its ability to address the needs of customers requiring a few oligonucleotides on a regular basis and those customers that require very large numbers of oligonucleotides on a less frequent basis. We will also respond to your special customisation needs – personally.
 

Features and Benefits

• Free Bio-RP purification: Near HPLC purity at a standard oligo price
• Quick turnaround time: < 3.5 business days from order to bench in most cases
• Broad range of modifications available: If you don’t see it, just ask, we can probably do it
• Competitive pricing: Great value for your research dollar

Why is Bio-RP purification better than desalting?

Most oligos sold are merely deprotected/desalted. The process of deprotection/desalting only partially purifies the oligo and leaves behind many impurities* and failure products (truncated oligos that will not amplify and that may interfere with certain applications). These can lead to artificially inflated OD readings that increase with oligo length.
Bio-RP purification removes these contaminants and failure products, resulting in an oligo that demonstrates near HPLC purity. The advantage to you is that our product contains only quality oligos minus impurities that may inhibit some PCR reactions and that potentially skew qPCR reactions.
We deliver the same amount of active oligo as our competitors; they just have more “other” artificially inflating their OD.
To demonstrate this, plates of Oligos of various length were tested for concentration in 2 steps:
1) following a deprotection/desalting step only, and 2) following Bio-RP purification. (Fig. 1)

The results of these tests show that the amount of failed sequences and impurities* increases with length (as expected), but the failure product is not removed by deprotection/desalting alone. This results in inflated OD readings for oligos depending on oligo length.
(Table 1)

Figure 1. Plates of Oligos of various length were tested for concentration following desalting and then by Bio-RP cartridge purification.
Note inflated OD readings when deprotection/desalting alone is used.
 

Table 1. Average OD inflation for oligos of various length when protection/desalting alone is employed v. Bio-RP purified product.
Note for longer oligos; up to 77% of product does not consist of functional oligo.

*Impurities in desalted oligos may include: Acetonitrile, Pyridine, Iodine,
Ethylthiotetrazole, Dichloromethane, Acetic acid, Acrylonitrile, Benzamide and Isobutyramide.

PAGE Oligo Purification

PAGE Oligo purification is recommended for Oligos between 50-100 bases. This results in yields with purity levels exceeding 95%. Subsequently yield obtained is lower than when other purification techniques are applied.

HPLC Oligo Purification

HPLC purification can be used for both modified and unmodified oligos up to 50 bases in length. Our HPLC oligo purification always begins with a trityl-on purification, and is often followed by a second full gradient trityl-off HPLC. In some cases, the initial trityl-on purification is entirely sufficient and a cartridge is utilized in the subsequent trityl-off oligo processing. This RP-HPLC purification process can result in yields of highly pure material, especially when the oligonucleotide is not complex. When synthesizing compounds that require post-synthesis modification such as dye labeling, we will apply an additional RP-HPLC oligo purification.

AccuOligo® prevents oligo dislodgment and potential loss during packing, shipping and opening through the addition of a patented adhesive to the tube during the production process. Even 96-deep well plate format orders have AccuOligo® technology applied, preventing cross-contamination from occurring during use.
(Patent application. Reg. Number 10-0777249 )

The adhesive does not affect PCR, Sequencing, restriction digests or other experimental methods
Prevention of dislodgment and loss of oligos during production, packaging and shipping
Prevents loss due to oligo "flaking" and tube cap attachment loss
Prevents cross-contamination when using plate formats
Quantified oligos made to order

Bioneer's in-house developed Bio-RP purification system.

Bioneer's R&D team has developed an RP resin for purification, as a superior alternative to standard desalting/deprotection as offered by our competitors. This delivers near-HPLC quality and is provided Free of Charge.
 

Contamination prevention through clean room production.

All oligos manufactured at Bioneer are produced within clean room facilities, ensuring that we provide nuclease/nucleic acid-free product to our customers.
 

Molecular weight-level quality control with accurate MALDI- TOF QC.

Every oligo produced is QC'd using MALDI-TOF and 96-well CE Q.C systems.
A comprehensive QC report is shipped with each oligo to ensure confidence and to ensure that our customers receive only the highest quality products for use in their research.

Ordering Information

Standard Custom
High Throughput
Modified
Dual Labeled Probes
Extendamers™
Large Scale

Bioneer's core business has always been the manufacture of DNA and RNA in conventional and breakthrough ways. When RNA interference (RNAi) was discovered, Bioneer was there to help provide researchers with the tools they need to accomplish their experiments. Bioneer is currently the market leader in RNAi technology in Asia, and is launching in North America this year. Our RNAi portfolio consists of six areas:

• Predesigned siRNA libraries
• PCR primers for knockdown validation
• Human validated siRNA libraries
• Human siRNA library sets and subsets
• Control siRNAs
• siRNA design and synthesis

All Bioneer's siRNAs are designed using our Turbo si-Designer software. This software identifies highly effective siRNA target sites with remarkably high knockdown rates. Critical parameters including base composition, thermodynamic instability and base preference are all considered in the design algorithm. siRNAs spanning SNP sites are removed and finally, non-specific siRNAs are eliminated following homology searching by BLAST and Smith-Waterman algorithms. The result is an siRNA with extraordinary knockdown efficiency and minimal off-target effects.We are so confident in our software that we have the best guarantee in the industry – Buy three of our AccuTarget™ siRNAs, and if 2 of them don’t have knockdown rates of at least 80%, we will replace them free of charge!

The Bioneer 80% Guarantee

When purchasing 3 siRNAs for the same gene, Bioneer guarantees at least an 80% reduction in the target mRNA level for 2 of the 3 siRNAs. If there is not a > 80% reduction in the mRNA level of the target gene, Bioneer will supply 2 siRNAs free of charge.*

*Bioneer reserves the right to request supporting data inclusive of:
1. Transfection efficiency data: NC (AccuTarget™ Fluorescein-labeled Negative Control) and siRNA concentration at 100 nM
2. siRNA knockdown efficiency data (mRNA level) : PC (AccuTarget™ GAPDH/GFP/Luciferase siRNA) and NC (AccuTarget™ Negative Control)
 

Features and Benefits

• Performance Guarantee: When purchasing 3 siRNAs for the same gene, Bioneer guarantees at least an 80% reduction* in the target mRNA level for 2 of the three siRNAs.

• Purification: For your more demanding applications, Bioneer's automated HPLC and Bio-RP purification methods ensure high quality, high- throughput siRNAs at an affordable price.

• Affordable pricing: Bioneer provides a variety of high quality siRNA products at an affordable price.

• Synthesis and QC: Bioneer siRNAs are produced in clean room facility by fully automated high-throughput siRNA production system. Bioneer siRNA products are assessed by MALDI-TOF Mass spectrometry analysis. Mass spec data is provided with every siRNA. Additionally, siRNAs are tested by gel electrophoresis to verify that both RNA strands annealed properly.

Until recently, gene knockdown or knockout technologies, such as antisense, ribozyme, and gene knockouts were used to perform loss-of-function studies. However, the post-genomics era calls for high-throughput gene function studies which the former technologies were unable to answer due to poor reproducibility, high cost, and excessive time to results. The advent of siRNA technology has opened up many new possibilities in the field of gene suppression.
 

siRNA Mechanism

siRNA is the term for 20 - 25-base pair RNA duplexes, where the two terminal 3'-nucleotides are unpaired (3' overhang). When siRNAs are introduced into cells they combine with a protein complex called the RNA-induced silencing complex (RISC) and are unwound by a helicase. The RISC complex containing single stranded RNA complementary to the target mRNA then recognizes and binds to the target mRNA. After binding the mRNA, the argonaute protein Ago2 cleaves it and complete degradation of the target mRNA is carried out by ribonuclease activity (as a result of the lack of protection by 5' caps or poly (A) tails). This exciting technology is one of the most effective methods for the silencing of specific target genes and is a must for gene function validation studies, drug target validation, and for gene therapy studies. siRNA has the following advantages over other RNAi technology:

• Reduced time and costs: Less screening is needed to obtain highly effective siRNA.
• High efficacy at lower concentration: lower concentrations provide effective gene silencing and minimizes off target effects
• Specificity: siRNA is a highly specific target knockout mechanism based on the natural biological mechanisms of RNAi


Bioneer manufactures high quality and cost-effective siRNA. Every siRNA is produced in an automated high-throughput RNA production system under clean room conditions and undergo rigorous QC tests.

All Bioneer siRNAs are provided as double-stranded siRNA. Each sense siRNA and an antisense RNA are QC'ed by MALDI-TOF. Every annealed siRNA is then QC-tested using PAGE to confirm proper annealing.


Figure 1. MALDI-TOF mass spectrometry analysis of a custom siRNA. All siRNAs are processed by MALDI-TOF mass spectrometry to ensure its quality.


Figure 2. PAGE data of double-stranded custom siRNA. Complementary single-stranded RNA strands were hybridized to form siRNA duplex and analyzed by 15% non-denaturing PAGE.
SS: single-stranded RNA
DS: double-stranded siRNA


Figure 3. Confocal microscopic image of HeLa cells transfected with FITC-labeled negative control siRNA (Cat No.: SN-1021). The fluorescent cells indicate that the HeLa cells were successfully transfected with the siRNA.


Figure 4. Effects of Human GAPDH Positive Control siRNA. HeLa cells were transfected separately with AccuTarget Human GAPDH Positive Control and Negative Control siRNA using Lipofectamine 2000 (Invitrogen) at a final concentration of 100 nM. Total cellular RNA was isolated from cells 24 hours after transfection and subjected to Northern blot and Real-Time PCR analysis. As can be seen from Fig. 1-B, about 3% GAPDH mRNA remained.


Notice to Purchaser

All siRNA Products: For Research Use Only. Not For Use in Diagnostic Procedures.

Limited License

This product is licensed under European Patents 1144623, 121945 and foreign equivalents from Alnylam Pharmaceuticals, Inc., Cambridge, USA and is provided only for use in academic and commercial research whose purpose is to elucidate gene function, including research to validate potential gene products and pathways for drug discovery and development and to screen non-siRNA based compounds (but excluding the evaluation or characterization of this product as the potential basis for a siRNA-based drug) and not for any other commercial purposes. Information about licenses for commercial use (including discovery and development of siRNA-based drugs) is available from Alnylam Pharmaceuticals, Inc., 300 Third Street, Cambridge, MA 02142, USA.

This product is sold for research use only and is not to be administered to humans or used for medical diagnostics. Buyer acknowledges and agrees that all intellectual property rights in the products (including, without limitation, the siRNA sequences used to create such products) and in any Bioneer technology, intellectual property and know-how used to make or useful for the manufacture or use of the products will at all times remain vested in Bioneer (other than any ownership interest that buyer may have in non-public proprietary target genes supplied by buyer to Bioneer in connection with custom products).

Trademark: AccuTarget is a trademark of Bioneer Corporation.

Q1. How is siRNA synthesized?

Q2. How do I order pre-designed siRNAs?

Q3. When can I expect delivery?

Q4. Up to how many bases can Bioneer synthesize?

Q5. What form will my order be in?

Q6. I want to conduct an in vitro experiment. What scale should I choose? What purification should I select?

Q7. Can Bioneer synthesize chimeric RNA?

Q8. I would like to order over four (4) siRNA candidates for a single gene. How do I order?

Q9. The Genome-wide predesigned siRNA didn't work like I expected. What do I do?

Q10. Are phosphate groups present on the 5' or 3' ends of the synthesized siRNA?

Modified siRNA

Q1. What types of modified siRNAs are there?

Handling and Storage

Q1. How to I store my siRNAs and how long do they keep?

Q2. How do I reconstitute my siRNA?

Q3. How to I store my fluorescent dye modified siRNA?

Purification

Q1. I would like to know how the synthesized siRNA is purified.

Quantification and Concentration

Q1. What does the O.D. value mean?

Q2. How do I calculate the amount of siRNA from the O.D. value?

Q3. If my 18-mer siRNA (3G, 4C, 5A, 6U) O.D. value was 0.7, then how much siRNA do I have?

Q4. How do I calculate the molecular weight of the synthesized siRNA molecule?

Q5. Can I know how many ng the synthesized product is?

Q6. How do I reach a target concentration?

Q7. How do I convert between morlarity and moles?

Experiments

Q1. What are some precautions for a siRNA experiment?

Q2. This is my first siRNA experiment. How do I set my experimental conditions?

Q3. To what cell density should I culture my cells before siRNA transfection?

Q4. What concentration of siRNA should I use for transfection?

Q5. How do I use 10 nmole of siRNA to transfect cells at 100 nM?

Q6. What transfection reagent do I use?

Q7. How do I verify my siRNA transfection efficiency?

Q8. Is there a way to know my siRNA's knockdown efficiency beforehand?

The predicted siRNA knockdown efficiency can be seen at the target gene search screen by clicking the siRNA Number. Validated siRNA efficiencies will be presented in blue, while predesigned siRNA efficiencies will be displayed in grey.

Q9. What controls are used for a siRNA experiment?

The AccuTarget Negative Control siRNA is a non-targeting siRNA that has low sequence homology to all known Human, Rat and Mouse sequences. Therefore, it can be used as a convenient negative control for all Human, Rat and Mouse siRNA experiments. The AccuTarget™ Positive Control siRNAs (human) shows great knockdown efficiency for the target gene. We target GAPDH as an endogenous control gene, and also have positive control siRNAs for reporter systems such as GFP and Luciferase.

Q10. How do I verify the siRNA knockdown efficiency?

The siRNA knockdown efficiency can be verified through various techniques including qPCR, Northern Blot, Western Blot etc.

Q11. What are AccuTarget Genome-wide Predesigned siRNAs?

These siRNAs were predesigned for your convenience using our Turbo si-Designer, a siRNA design algorithm co-developed by us and KRIBB (Korea Research Institute of Bioscience & Biotechnology). Using this revolutionary algorithm, we have constructed a high-knockdown efficiency AccuTarget Genome-wide Predesigned siRNA database. As of this writing, our website contains predesigned siRNAs for Human (17,842 genes), Mouse (17,117 genes), and Rat (9,392 genes).

Q12. What are AccuTarget validated siRNAs?

After synthesizing siRNAs using our Turbo si-Designer, a siRNA design algorithm co-developed by us and KRIBB (Korea Research Institute of Bioscience & Biotechnology), AccuTarget Validated siRNAs have been measured for knockdown efficiency using qPCR. We verified that each siRNA shows at least a 70% knockdown efficiency. AccuTarget Validated siRNAs can be ordered with their corresponding real-time qPCR primer sets. Also, if you order 10 or more siRNAs, they will be delivered in flexible siRNA Library (tube or plate type) format. 

Q13. Can I order siRNAs argeted for a specific gene?

You may order siRNAs for the target gene of your choice by entering the gene name or Accession number at our website and selecting one of the listed siRNA candidates. You may also order custom designed siRNA if your model organism is not Human, Mouse or Rat.

Q14. What are precautions for handling siRNA?

- Wear gloves and a mask when handling siRNA.
- Always use RNase free tubes and pipette tips.
- Only use DEPC treated water when diluting siRNA.
- Store fluorescence-labeled siRNA in a dark location.
 
 

With the latest technology and years of experience designing and manufacturing products for protein synthesis and purification, Bioneer is pleased to showcase our Accu- and Exi- series products for protein science. These products are designed to simplify your protein work in a cost-effective way.
 

Features and Benefits

ExiProgen™/ExiProgen™ EC1 Protein Synthesis Kit

Fully automatic: DNA-in-Protein-out:
ExiProgen™ EC1 Kit and AccuRapid™ Cell-Free Protein Expression Kit require only the addition of template DNA which can be in to form of an expression vector such as pBIVT or a PCR product as can be made with the ExiProgen ProXpress PCR Template Kit.

Parallel Process
Obtain from 1 to 16 different proteins in a single run.

High performance
All systems allows for synthesis of proteins that are toxic to in vivo expression systems.

Reproducibility
The only hands-on step is the addition of template DNA, maximizing reproducibility.

Fast procedure
The in vitro (cell-free) protein synthesis system does not require time-consuming cell culture processes.
 

AccuRapid™ Cell-Free Protein Expression Kit/ ExiPrep™ & AccuPrep® His-tagged Protein Purification Kit

High-speed
AccuRapid™ Cell-Free Protein Expression Kit reactions are carried out in just 3 hours. His-Tag purification can be accomplished in 30 minutes!

Simplicity
The AccuRapid™ Cell-Free Protein Expression Kit contains all necessary components for transcription and translation. Simply add template and incubate!

Flexible
The AccuRapid™ Cell-Free Protein Expression Kit can synthesize proteins from various DNA templates.

SSMB (Spherical Shape Magnetic Beads) for His-Tag purification
SSMB beads have increased surface area by virtue of their size and eliminate the carryover problems of rough surfaced magnetic particles.

AccuRapid™ Cell-Free Protein Expression Kit
This product is an adaptation of a cell-free protein expression system.
It includes all necessary components for successful in vitro protein expression. A total of twenty-four (24) 45uL volume reactions are possible with this product.

AccuPrep® is-tagged Protein Purification Kit
This kit is designed for the convenient purification of Histidine-tagged proteins from E. coli lysate.
Purification is based on Ni-NTA magnetic beads for magnetic or centrifugal purification.

2. Exi- series: ExiProgen series products are designed for Bioneer's automated systems (ExiProgen™).

ExiProgen™ EC1 Protein Synthesis kit
This product is designed to yield world-class protein synthesis performance when combined with the world’s first automated protein synthesis and nucleic acid purification system ExiProgen™. Up to 16 different proteins can be expressed and purified simultaneously within 6 hours of adding template DNA, and automation means that the results will be reproducible run after run.

ExiProgen™ His-tagged Protein Purification Kit
This product is designed for the purification of up to 16 His-tagged proteins simultaneously within 30 minutes with excellent reproducibility from automation

Bioneer offers a wide range of room temperature stable, ready-to-load DNA ladders as well as molecular weight markers for electrophoresis. Our ladders contain bands ranging from 10 bp to 10,000 bp, and our markers have bands from 125 bp to 23,000 bp.
The number of fragments generated by each DNA marker, as well as the specific fragment sizes for each of the conventional markers can be found in our selection guide, or on the specific product page.

Features and Benefits

• Convenient: Both ladders and markers provided ready-to-load
• Robust stability: Stable for 4 months at room temperature
• Broad range of products available: You are sure to find one that meets your needs
• Manufactured with strict control standards: Uniform and sharp band intensities
• Competitive pricing: Great value for your research dollar

The world's first S. pombe Genome-wide Deletion Mutant Library
This genome-wide S. pombe deletion collection covers more than 98% of genome consisting of 4,914 genes. As each deletion strain has its distinct tags, functional analysis can be performed both in a gene-specific manner and a genome-wide scale. Two different sets of mutant collections are available: h+ haploids and h+/ h+ diploids.

S. cerevisiae VN-Fusion Library

Powerful Tool for Protein-Protein Interaction Analysis
The yeast Saccharomyces cerevisiae(S. cerevisiae) is a recognized simple eukaryote model system which genome can be easily manipulated. The S. cerevisiae VN-Fusion Library was created by Dr. Won-Ki Huh of Seoul National University (Korea). The VN-Fusion Library consists of 5,809 VN-tagged Open Reading Frames (ORFs) covering 93% of the yeast proteome.

Features & Benefits of GPScreen™ Services

• World's unique & innovative technology
• Identify any classes of drug targets in genome through genome-wide screening
• Live cell-based screening
• High throughput screening
• Rapid; average 20 days for genome-wide screening
• Accurate, precise & cost-effective
• Drug target validation services available in human cells (optional !)
• Applicable to drug repositioning, natural drug target discovery, toxicity profiling

Ⅰ. Introduction

Precise Drug Target Identification is a first step for improving efficacy, tracing and avoiding side-effects as well as understanding the mode-of-actions of drug candidates. However, until now, effective systematic approaches for the precise drug target identification at genome levels have not been commercially available. Using S. pombe genome-wide heterozygous deletion mutant library, Bioneer has developed a high-throughput genome-wide drug target screening service system (GPScreen™) and made it commercially available for your drug discovery needs. GPScreen™ screening service covers a broad spectrum of drug candidates in whole disease areas from cancers & metabolic diseases to neglected & rare diseases and, therefore, will provide the total solutions for an efficient drug discovery through providing clear-cut answers to problems such as drug-target(s) and toxicity as well as mode-of-actions of drug candidates.


 

“Bioneer’s GPScreen™ using S. pombe genome-wide heterozygous deletion mutant library has enabled genome-wide screening for drug target identification”

GPScreen™ is based on Bioneer’s unique S. pombe genome-wide deletion mutant library, which was developed by Bioneer and the Korea Research Institute of Bioscience & Biotechnology (KRIBB) in collaboration with Dr. Paul Nurse of Cancer Research Center UK (Nat. Biotech, 28, 617–623 (2010)). It can be used for genetic and chemical screening such as drug target identification, gene expression profiling and synthetic lethal profiling.

This mutant library comprised of a total of 4,840 heterozygous diploid deletion mutants representing 98.5% genome coverage and one copy of a specific gene in each mutant was deleted by homologous recombination. GPScreen™ exclusively offers S. pombe deletion mutant library as a powerful tool for large-scale genetic functional analysis, identification and verification research of drug targets and for integrated systems research of cell function.


Ⅱ. Principle of GPScreen™

"Drug-induced Haploinsufficiency (DIH)" is a situation in which a strain with a heterozygous gene deletion (producing target proteins to about half of normal levels), result in cells sensitive to a specific drug. DIH occurs when a drug or drug candidate acts on a mutant strain in which the level of target gene protein is lowered from normal to half level, and is considered to be a valuable tool for drug target identification. Previously, a number of reports have provided identifications of drug targets using DIH in budding yeast S. cerevisiae (Baetz K et al., 2004; Lum et al., 2004). However, fission yeast S. pombe is considered a better model of mammalian cells since its cell cycle regulation is closer to that of mammalian cells than that of S. cerevisiae

“GPScreen™ utilizes DIH in the fission yeast S. pombe genome-wide heterozygous deletion mutant library to identify the drug target(s).

References

• Baetz K et al. Yeast genome-wide drug-induced haploinsufficiency screen to determine drug mode of action. PNAS. 2004
[More information]

• Lum PY et al. Discovering modes of action for therapeutic compounds using a genome-wide screen of yeast heterozygotes.
Cell. 2004
[More information]

Ⅲ. Performance of GPScreen™

GPScreen™ is a best-in class genome-wide drug target identification system that is exclusively offered by Bioneer Inc. It allows for the clear-cut identification of molecular targets of drugs/ drug candidates (A representative is below).

Figure 1. Drug Target Identification using GPScreen™.
In the presence of cytochalasin A (Fig. 1A), a heterozygous deletion mutant of act1 shows decreased growth as a result of a decrease in “functional” Act1 protein. act1 was the only gene in the genome-wide screen to show this effect, demonstrating that act1 is a target of cytochalasin A (Fig. 1B).


Ⅳ. Applications of GPScreen™

Toxicity profiling and Drug Prioritization
Drug Repositioning and Enhancement
Natural Drug Target Discovery
Chemogenomic profiling

Toxicity profiling using GPScreen™ at early stage of drug development would accommodate 'Drug Prioritization' among drug candidates very efficiently, thereby reducing time and money dramatically in the later stage of drug discovery.


V. Flow chart of GPScreen™ Service

ORTHentication Services (Ortholog-Authentication services) provide an siRNA-based gene susceptibility test for human orthologs identified through GPScreen™. This allows for validation of the drug targets in human cells. This is done via a method that is similar to Haploinsufficiency screens in yeast (See our GPScreen™): the cells are exposed to the drug and a GI₅₀ is determined for wild type cells. Then they are exposed to the same level of drug after siRNA treatment and subsequent knockdown of the target gene. If the gene in question is being targeted by the drug candidate, then the knockdown of the gene would have a further deleterious effect on cell growth – clearly validating the Human ortholog as a drug target.

Target validation services include

• Discovering human orthologs of drug target genes identified through GPScreen™ via rigorous bioinformatics analysis
• Design and synthesis of siRNAs targeting the human orthologs using our exclusive genome-wide human siRNA library (AccuTarget Genome-wide Predesigned siRNA library)
• Validation of the drug target in human cells by co-treatment of siRNA targeting the corresponding human ortholog (direct mimicking of haploinsufficiency of S. pombe system) followed by the drug treatment
• Cell biology analysis including proliferation assays in the presence of siRNA and drug; analysis of phenotypic outcome and mode-of-action of the drug

ORTHentication Services are especially well suited for rapid drug repositioning and allow you to fast-track to clinical trial using drug candidates whose toxicity and efficacy tests have been previously completed.

in vivo Validation of ORTHentic (Ortholog-Authentic) genes

Once positively identified, the Human Orthologs can be further validated in an in vivo mouse model. To achieve this, the mouse homolog of the human gene is pulled from our genome-wide mouse siRNA library. This gene is then synthesized into our novel SAMiRNA (Self-Assembled Micelle siRNA) prodrug using a proprietary synthesis method. SAMiRNA is a novel prodrug that is stable in vivo for several days and can be targeted to cell types through targeting molecules on its surface. SAMiRNA also naturally targets Tumors and cancerous tissue through enhanced permeability and retention effects. It has been demonstrated to have no detectable toxicity at therapeutic levels and is the best vehicle for in vivo siRNA delivery available.

 

SAMiRNA (Self-Assembled-Micelle-interfering-RNA)

One of the major hurdles for the development of RNA interference (RNAi)-based therapeutics is siRNA delivery. SAMiRNA (Self-Assembled-Micelle-interfering-RNA) is a novel class of RNAi molecule developed by Bioneer. The therapeutic potential of SAMiRNA is highlighted by virtue of its negligible toxicity and superb in vivo serum stability as well as its target gene silencing efficacy. These data indicate SAMiRNA’s exceptional therapeutic potential as an RNAi platform for a variety of diseases. Development of SAMiRNA system combines Bioneer’s extensive nucleic acid chemistry and pre-clinical research with our nucleic acid manufacturing and development expertise, which has been providing top-quality DNA oligos and siRNAs worldwide for close to 20 years.

Delivery challenges overcome by SAMiRNA technology

Bioneer’s SAMiRNA Custom Services provide a wide range of RNAi research services to companies and research institutions worldwide. We offer a complete solution for RNAi (gene silencing) studies and drug discovery. The company’s research and technical support teams ensure the top-quality products and services to meet the unique needs of clients. Our world-class teams have extensive nucleic acid therapeutic manufacturing and development expertise.

Our SAMiRNA Custom Services include:

Custom siRNA design/synthesis (also offering genome-wide pre-designed mouse and human siRNA library)

in vitro screen/validation of siRNAs of choice

SAMiRNA nanoparticle synthesis

in vivo siRNA delivery and biodistribution/efficacy test using murine models

Quantitation of functional knockdown of target gene by real time qRT-PCR

And much more

Features and Benefits

Novel, self-assembling siRNA nanoparticles form with a protective PEG coat and optimal nano-scale size for selective localization in either vascularized tumors through EPR effects, or to targeted tissues through the addition of targeting moieties to the surface of the SAMiRNA.

Novel patent-pending Self-Assembling, siRNA nanoparticles (Figure 1) form with a protective PEG coat and optimal nano-scale size for selective localization in either vascularized tumors through an Enhanced Permeability and Retention (EPR) effects, or to targeted tissues through the addition of targeting moieties to the surface of the SAMiRNA.

• siRNA Prodrug: highly stable in circulation and siRNA is released and active only when metabolized within the target cells
• Combined with various targeting moiety, SAMiRNA nanoparticles can target various target organs of interest, for example, liver.
• Suitable for studying in vivo function of tumor related genes in clinically relevant manner (Figure 2 and 3)
• The most cost effective way to prioritize promising gene targets for drug discovery and validate candidate RNAi therapeutics in vivo
• No detectable toxicity or cytokine induction, minimizing off-target effects

A new class of single molecule-based self-assembling Nanoparticles

Figure 1. Structure of SAMiRNA Nanoparticle.
(A) Schematic diagram of SAMiRNA (B) Cryo-TEM images of SAM-siRNA Nanoparticles (scale bar = 100 nm).

Figure 2. in vivo silencing of target mRNA by SAMiRNA Nanoparticels (NPs).
(A) Tumor bearing mice were intravenously injected with saline (PBS), SAMiRNA, and tumor-targeting folic acid (FA)-conjugated SAMiRNA Nanoparticles at a single 5mg/kg dose. Mice were sacrificed at denoted time points and targeted survivin mRNA levels in isolated tumor cell mass were subsequently measured by Real-Time PCR. (B) Tumor bearing mice were intravenously injected with saline (PBS) and SAMiRNA NP at a single 1 or 5mg/kg dose. Mice were sacrificed at 72 hr postinjection and targeted survivin mRNA levels in isolated tumor cell mass were subsequently measured using Real-Time qPCR.

Tumor-specific targeting of SAMiRNA NPs in mouse cancer models

This service was designed to meet your specific needs fast (results in only 2-3 weeks) at extremely attractive prices. GPScreen™ services is provided with not only genome-wide screening (using 4840 full mutant strains) or as essential gene screening (using 1259 essential gene mutant strains). Furthermore, we can also provide other screening services on specified functional gene groups which are involved in various signaling pathways and/or disease areas (e.g. Cell cycle regulation/cancers) in accordance with your specific needs: By screening only a specified functional subset of the library you can use this service at a reduced price as described below.



 

To place an order, please follow these steps:
• Download the ordering form. [Download] Note that you will need to fill out one Worksheet/drug compound being tested.
• Complete the order form with your information, product, drug and other relevant info.
• E-mail the completed order form to contact@bioneer.com.au
• Bioneer will confirm order and send service information (service price & period) via e-mail.
• The service can be started the same day the compounds are received.

Ⅱ. How to send us your compounds

* Delivery address:
S. pombe Research Team, Bioneer Corporation
8-11, Munpyeongseoro, Daedeok-gu, Daejeon 306-220, Republic of Korea

(When shipping your compounds, please write your name and organization on your box.)

Ⅲ. Contact Us

For order inquiries and status, please email us at contact@bioneer.   com  .au

Ⅳ. GPScreen™ Service

GPScreen™ Service provides various screening services to meet the specific needs of customers. Therefore, you can make the best choice among the various services below. After considering the issues and problems of your drug candidates, choose any service you need.

1) Primary Test Service for Determination of Growth-inhibitory Activity (GI₅₀) in Wild Type S. pombe

Primary Test Service will be performed to all compounds requested in advance of GPScreen™

* Price of primary test will be dramatically discounted when the number of compounds is ≥ 2.

2) GPScreen™ Service using S. pombe Genome-wide Mutant Set

3) GPScreen™ Service using S. pombe Essential gene Mutant Set

 4) GPScreen™ Service using KOG analysis-based Functional Group Subsets

5) GPScreen™ Service using Human Disease-related Subsets

Ⅴ. SAMiRNA in vivo Drug Target Screening

Bioneer’s SAMiRNA Services will provide a rapid e-mail quotation (contact@bioneer.com.au),
comprehensive service packages, and highly competitive prices.
Please let us know the details of your project, so that we can provide you with an accurate quotation and timeline estimate.

ExiProgen™ from Bioneer

The first instrument for Automatic Protein biosynthesis, DNA/RNA purification

ExiProgen™ is unique instrument for automated protein biosynthesis in vitro. It features the ability to express 16 different proteins simultaneously only using recombinant plasmid. Also it is able to purify His- tagged proteins automatically without any additional column work. As well as His-tagged proteins, DNA/RNA from various biological samples such as whole blood, tissue, mammalian cells, bacteria, and plants can be purified and extracted with high quality. ExiProgen™ has advanced features such as an intuitive touch screen, UV sterilization, and Contamination Shield, each designed to provide an extra level of confidence in your results. ExiProgen™ is essential instrument for 21C Biotechnological laboratory

Exicycler™ 96 from Bioneer

Superior 5-color Real-Time Quantitative PCR system

Exicycler™ 96 is a superior 96-well PCR system designed for real-time qPCR applications demanding the highest performance. Proprietary Light Tunnel (LT) Technology mitigates the common problem of uneven light distribution across the 96-well plate, allowing for normalization of fluorescence signals from all five channels without the use of a reference dye. The instrument is also equipped with a thermal gradient function, allowing for convenient PCR condition optimization experiments. The analysis software contains several different modules for analyzing qPCR data including: Absolute Quantification, Relative Quantification, SNP genotyping, Existence/Nonexistence and melting curve analysis. With the various modules analyses such as viral load monitoring, gene expression analysis, SNP genotyping etc. can be performed. In vitro diagnostic software packages have also been developed (KFDA, CE-IVD certified versions), which allows the instrument to be used for molecular diagnostic purposes.

Bioneer Pacific - Product Ordering

Nov 2013: Victor Chang Cardiac Research Institute acquires ExiProgen^TM                        
Bioneer Pacific is pleased to announce that the Victor Chang Cardiac Research Institute (VCCRI) in Sydney has elected to acquire the ExiProgen™ Protein Synthesis and Purification system as a system platform critical for the acceleration of its protein research.
 
The following news release can be found at the Victor Chang Cardiac Research Institute website.
http://www.victorchang.edu.au/home/news-events/news/news-detail/?news_name=new-method-rapid-production-protein
 

                                                  NEW METHOD FOR RAPID PRODUCTION OF PROTEINS 

A new instrument, the ExiProgen^TM from Bioneer, is allowing scientists at the Victor Chang Cardiac Research Institute to rapidly compare normal proteins, the powerhouse of the body’s cells, with those found in diseases.
The ExiProgen^TM enables scientists to manufacture and purify a protein within a cell-free in-vitro system, rather than using the traditional method, which involves expressing it from live cells.
Dr Romaric Bouveret said: “This instrument is the first of its kind in Australia. By using it to look at the structure of normal proteins, we hope we can then work backwards to better understand the effect of mutant proteins, responsible for different cardiovascular conditions including congenital heart disease.
“It allows us to look quickly at lots of different conditions which might affect the production and stability of proteins, including temperature.
“The machine has also been designed to enable the rapid purification of total DNA and RNA from cells, which means we can obtain high yields of ultra pure material very quickly.  Although we haven’t explored these capabilities yet; we are very excited about doing so, and benefitting much more from the machine in the future.”
The Victor Chang Cardiac Research Institute was able to purchase the ExiProgen^TM thanks to the generosity of Mrs Mimi Wong.
 
The Victor Chang Cardiac Research Institute is an independent, not-for-profit research facility, dedicated to the memory of cardiac surgeon Dr Victor Chang and his passionate belief in the power of discovery. Founded in 1994, the Institute has grown to become a world-class cardiac research and research training facility.
The ExiProgen™ system has been developed by Bioneer Corporation, a KOSDAQ listed company based in Daejeon, South Korea.

 

Feb 2013: Bernice Tan joins Bioneer Pacific
Bernice Tan has joined Bioneer Pacific as a Sales & Applications Specialist. Bernice studied at the University of New South Wales and comes to us having recently completed her PhD at the Garvan Institute of Medical Research. She has considerable depth and experience in many aspects of molecular biology, including protocol optimisation and troubleshooting which is sure to add substantial value for our customers when considering and selecting products from the Bioneer Pacific product range for their research. Welcome Bernice.




Feb 2013: Bioneer Pacific at the Lorne Genome Conference
Bioneer Pacific will be participating at the 34^th Annual Lorne Genome Conference at the Mantra Lorne Resort Feb 17^th – 19^th.
We will be showcasing the ExiProgen™ ‘cell-free’ protein synthesis and purification system in addition to our comprehensive range of Amplification kits and Enzymes.
Please drop by to see us at booth #4 to meet us and to pick up your 10% discount Coupon Code in celebration of the launch of our new Australian website. The Coupon Code can be used right across the product range and includes online ordering of Oligos.

 

The AccuPower® Taq PCR PreMix is a convenient lyophilized PCR master mix containing Taq DNA polymerase, dNTPs, reaction buffer, tracking dye, and patented stabilizer and is aliquoted in 8-strip PCR tubes. The premix retains its activity for over a month at room temperature and is stable for two years in -20°C freezer. AccuPower® Taq PCR PreMix is available with or without tracking dye, depending on your application. If purchased with tracking dye, reactions can be loaded on agarose gels without adding loading buffer.

 Features and Benefits

• Flexible: Taq provides accurate amplification of standard and highly suitable for all PCR applications.
•  Easy to use: All reaction components required for PCR, including thermostable DNA polymerase and dNTPs are contained within each tube and in a lyophilized "PreMix" form. 
•  Reproducibility: Bioneer's strict quality controlled production system ensures that your results will be reproducible experiment after experiment.
•  Convenient: Just add template and primers and start your reaction. dNTPs, buffer and enzyme are provided
•  Stability: Stable at room temperature for a month and for 2 years in a -20°C freezer

Source: thermus aquatics
5' -> 3' exonuclease activity: Yes
3' -> 5' exonuclease activity: No
3' – A Overhang: Yes

Application

- Routine PCR
- Primer extension
- TA cloning
- Gene sequencing

Experimental data


Figure 1. Comparison of PCR amplification efficiency between AccuPower® Taq PCR PreMix from Bioneer and other suppliers' PCR master mix.

The cycling conditions for AccuPower® Taq PCR PreMix were 95°C for 5 min, 35 cycles of 95°C for 20 sec, 55°C for 20 sec, and 72°C for 30 sec. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
Target Gene : human myc.
Lane M : 100 bp DNA Ladder (Bioneer, Cat. No. D-1030)
Lane 1 : 10 ng of human total cDNA
Lane 2 : 1 ng of human total cDNA
Lane 3 : 100 pg of human total cDNA
Lane 4 : 10 pg of human total cDNA
 

 
Figure 2. Comparison of PCR amplification efficiency between AccuPower® Taq PCR PreMix from Bioneer and other suppliers' PCR master mix.

The cycling conditions for AccuPower® Taq PCR PreMix were 95°C for 5 min, 35 cycles of 95°C for 20 sec, 55°C for 20 sec, and 72°C for 30 sec. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
Target Gene: IRGC (Immunity-related GTPase family, cinema)
Lane M : 100 bp DNA Ladder (Bioneer, Cat. No. D-1030)
Lane 1 : 10 ng human genomic DNA
Lane 2 : 1 ng human genomic DNA
Lane 3 : 100 pg human genomic DNA
Lane 4 : 10 pg human genomic DNA
 

Figure 3. Comparison of PCR amplification of long targets between AccuPower® Taq PCR PreMix from Bioneer and other suppliers' PCR master mix.

The cycling conditions for AccuPower® Taq PCR PreMix were 95°C for 5 min, 30 cycles of 95°C for 20 sec, 65°C for 20 sec, and 68°C for 8 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
Lane M : 1 kb DNA Ladder (Bioneer, Cat. No. D-1040)
Lane 1 : 3 kb fragment (human tumor protein p53 gene)
Lane 2 : 4 kb fragment (human beta globin region)
Lane 3 : 4.5 kb fragment (human DNA cross-link repair 1A gene)
Lane 4 : 8 kb fragment (human hemoglobin epsilon 1 gene)

• AccuPower® Taq PCR MasterMix

MSDS
• AccuPower® Taq PCR PreMix

Brochure
• AccuPower® PreMix series - 2010 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

The polymerase chain reaction (PCR) can amplify a single molecule over a billionfold. Thus, even minuscule amounts of a contaminant can be amplified and lead to a false positive result. Such contaminants are often products from previous PCR amplifications (carry-over contamination). Therefore, researchers have developed methods to avoid such contamination AccuPower® PCR PreMix(with UDG) is a ready-to-use master mix containing all components, except primers, for the amplification and detection of DNA in PCR The master mix combines Top DNA polymerase with integrated UDG carryover prevention technology to provide optimal performance with a variety of PCR detection technologies

Features and Benefits

Prevention of carryover contamination
UDG and dUTP in the MasterMix prevent the reamplification of carryover PCR products between reactions. dUTP ensures that any amplified DNA will contain uracil, while UDG removes uracil residues from single- or double-stranded DNA, preventing dU-containing DNA from serving as template in future PCRs, A UDG incubation step(37°C,2min) before PCR cycling destroys any contaminating dU-containing product from previous reactions. UDG is then inactivated by the high temperatures during normal PCR cycling, thereby allowing the amplification of genuine target sequences




Easy-to-use
All reaction components required for PCR, including thermostable DNA polymerase and dNTPs are contained within each tube and in a lyophilized "PreMix" form. The user needs only to add template DNA, primers and water. Materials necessary for loading agarose gels for electrophoresis are also added in the reaction, negating the need to add loading dye after PCR is completed

Reproducibility
Each batch is produced under strict quality controls. Errors that commonly occur during mass production are eliminated during the individual packaging process. Bioneer’s current batch processing system allows for the production of more accurate and reproducible end-product yield

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: Yes
Fragment Size: ~ 10 kb

Application

Polymerase chin reaction (PCR)
Primer extension
TA cloning
Gene sequencing
Molecular Diagnosis

Experimental data

Sensitivity


Figure 1. Comparison of sensitivity between Accupower® PCR PreMix and Accupower® PCR PreMix(with UDG).
Sensitivity test was operated using serial diluted human genomic DNA. Reaction mixture was incubated at 37°C for 2min followed by 95°C for 5min, 30 cycles of 20sec at 95°C, 20sec at 55°C, 30sec at 72°C. Accupower® PCR PreMix(with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP. The amount of DNA used to test is represented below.
Lane 1: 10ng of Human genomic DNA Lane 2: 1ng of Human genomic DNA
Lane 3: 100pg of Human genomic DNA Lane 4: 10pg of Human genomic DNA
M: 100 bp DNA ladder (Bioneer Cat. No: D-1030-1)

MSDS
• MSDS_AccuPower® PCR PreMix(with UDG)

Brochure
• AccuPower® PreMix series - 2010 Brochure

Quality Assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

Hot Start PCR master mix, unique Hot Start method for DNA amplification

AccuPower® HotStart PCR PreMix is a PCR master mix containing a thermostable DNA polymerase, thermostable pyrophosphatase, reaction buffer, dNTPs, tracking dye, and patented stabilizer in a ready to use hotstart PCR master mix.

Bioneer uses a unique enzyme-mediated HotStart PCR that provides robust and reliable results. Bioneer’s Top DNA Polymerase is inhibited at lower temperatures (< 70°C) by pyrophosphate. However, Top DNA Polymerase is rendered fully active at temperatures above 70°C via pyrophosphate hydrolysis with our thermostable pyrophosphatase. This prevents the formation of mis-primed products and primer-dimers during the reaction set up process resulting in improved PCR specificity. It is ideal for nucleic acid amplification reactions involving complex genomic or cDNA templates, very low copy targets, and multiplex reactions.

Features and Benefits

• Ease of use: Just add template and primers and start your PCR, dNTPs, buffer and enzyme are provided
• Stability: Stable at room temperature for a month and for 2 years in a -20°C freezer

•  HotStart: Unique enzyme mediated HotStart results in greater specificity and more robust reactions

Application

HotStart PCR up to 12 kb
PCR with complex genomic templates/low copy
Templates cDNA
Multiplex PCR reactions



Figure 1. Specificity comparison between AccuPower® PCR PreMix and AccuPower® HotStart PCR PreMix. 50 ng human genomic DNA was amplified into 2.5 - 3 kb fragments with the two products.

A: AccuPower PCR PreMix
B: AccuPower HotStart PCR PreMix
Lane M: 1 kb DNA Ladder (D-1040)
Lane 1: 2.5 kb product
Lane 2: 3.0 kb product



Figure 2. Specificity comparison of multiplex PCR with various HotStart PCR reagents. Each PCR mixture with 2 ng of human genomic DNA and 2 pairs of primers (p53 + p55 and p55 + p63) was incubated for 2 hours at 37°C prior to performing multiplex PCR.

M: 100 bp DNA Ladder (D-1030)
A: AccuPower® HotStart PCR PreMix
B: Control – AccuPower® PCR PreMix
C: Company T's antibody mediated HotStart Taq DNA polymerase (0.5 U)
D: Company A’s antibody mediated HotStart Taq DNA polymerase (1.0 U)

MSDS
• AccuPower® Hotstart PCR PreMix

Brochure
• AccuPower® PreMix series - 2010 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

Features and Benefits

• Optimized 5X buffer: For more full length fragments - within 10 minutes

• Full length cDNA: For genes up to 9 kb

• RNase, DNase and Proteinase-free: Ensures the integrity of your samples

5' to 3' exonuclease activity: No
3' to 5' exonuclease activity: No
3’ – A Overhang: No
Strand Displacement: Yes Fragment Size: Up to 9 kb

Application

First strand cDNA synthesis from RNA, RT-PCR, and qRT-PCR

Reagents Supplied

5 x Reaction Buffer: 50 mM Tris-HCl, 250 mM KCl, 10 mM MgCl₂, pH 8.1
100 mM DTT
dNTP Mix: 2.5 mM of each dNTP

Concentration

200 U/μl

Storage Conditions

50% glycerol containing 20 mM Tris-Cl (pH7.6), 150 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 0.1 % IGEPAL CA-630

Store Temperature

-20°C

Unit Definition

One unit is defined as the amount of enzyme required to incorporates 1 nmole of dTTP into acid-precipitable material in 10 minutes at 37°C using poly (A)oligo (dT) as template primer.

• M-MLV Reverse Transcriptase

MSDS

• M-MLV Rtase

• M-MLV Rtase dilution buffer

• M-MLV reaction buffer

• 100mM DTT

Brochure

• Enzymes 2010 Brochure

Quality Assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

• EC Directive 98/79/EC - certificate

Questions or Complaints
If you have any questions or complaints regarding the privacy or security of information you should contact the Privacy Officer, Bioneer Pacific on 1300 414 053 in the first instance.

Legal information regarding Copyrights and trademarks

AccuPower

Features and Benefits
 

• Long PCR:  ProFi Taq is especially effective in amplifying large genomic DNA fragments around 20 kb and amplifying Lambda DNA up to 30kb.
• Easy to use:  All reaction components required for PCR, including thermostable DNA polymerase and dNTPs are contained within each tube and in a lyophilized "PreMix" form. 
• Reproducibility:  Bioneer's strict quality controlled production system ensures that your results will be reproducible experiment after experiment.
• Convenient:  Just add template and primers and start your reaction. dNTPs, buffer and enzyme are provided
• Stability:  Stable at room temperature for a month and for 2 years in a -20°C freezer

5' to 3' exonuclease: Yes
3' to 5' exonuclease: Yes
3' – A Overhang: Yes
PCR product size: ~ 30kb

Application

- Primer extension
- long-range amplification from genomic DNA
- High amplification efficiency
- Excellent performance on difficult templates
- Amplification of low-copy targets
- High yield and high sensitivity PCR

Experimental data


Figure 1. Comparison of PCR amplification efficiency between AccuPower® ProFi Taq PCR PreMix from Bioneer and other suppliers' PCR master mix

The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5min, 30 cycles of 95°C for 20 sec, 55°C for 20 sec and 72°C for 30 sec. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
Target : human insulin receptor gene.
Lane M : 100 bp DNA Ladder(Bioneer, Cat. No. D-1030)
Lane 1 : 10 ng of human genomic DNA
Lane 2 : 1 ng of human genomic DNA
Lane 3 : 100 pg of human genomic DNA
Lane 4 : 10 pg of human genomic DNA


Figure 2. Comparison of PCR amplification efficiency between AccuPower® ProFi Taq PCR PreMix from Bioneer and other suppliers' PCR master mix

cDNA synthesized from 10-fold serial-diluted human total RNA from 10 ng to 10 pg using AccuPower® RocketScript™ Cycle RT PreMix. (Bioneer, Cat. No K-2201) wase used as a template for PCR amplification. The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5min, 33 cycles of 95°C for 20 sec, 55°C for 20 sec and 72°C for 30 sec. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
Target : human GAPDH gene.
Lane M : 100 bp DNA Ladder(Bioneer, Cat. No. D-1030)
Lane 1 : 10 ng of human total cDNA
Lane 2 : 1 ng of human total cDNA
Lane 3 : 100 pg of human total cDNA
Lane 4 : 10 pg of human total cDNA

The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5 min, 30 cycles of 95°C for 20 sec, 65°C for 20 sec and 68°C for 4 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
Lane M1 : Lambda/Hind III marker (Bioneer, Cat. No. D-1050)
Lane M2 : 1 kb DNA Ladder (Bioneer, Cat. No. D-1040)
Lane 1 : 2 kb fragment (human tumor protein p53 gene)
Lane 2 : 3 kb fragment (human tumor protein p53 gene)
Lane 3 : 4.5kb fragment (human DNA cross-link repair 1A gene)
Lane 4 : 8 kb fragment (human hemoglobin epsilon 1 gene)


Figure 4. Comparison of PCR amplification of long targets between AccuPower® ProFi Taq PCR PreMix from Bioneer and other suppliers' PCR master mix

The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5 min, 32 cycles of 95°C for 20 sec and 68°C for 15 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol. Human genomic DNA was used as a template for PCR amplification.
Lane M1 : Lambda/Hind III marker (Bioneer, Cat. No. D-1050)
Lane M2 : 1 kb DNA Ladder (Bioneer, Cat. No. D-1040)
Lane 1 : 11 kb fragment
Lane 2 : 13.5 kb fragment
Lane 3 : 17.6 kb fragment
Lane 4 : 21.4 kb fragment


Figure 5. Comparison of PCR amplification of long targets between AccuPower® ProFi Taq PCR PreMix from Bioneer and other suppliers' PCR master mix

The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5 min, 32 cycles of 95°C for 20 sec, 65°C for40 sec, and 68°C for 20 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol. Lambda DNA was used as a template for PCR amplification.
Lane M1 : Lambda/Hind III marker (Bioneer, Cat. No. D-1050)
Lane M2 : 1 kb DNA Ladder (Bioneer, Cat. No. D-1040)
Lane 1 : 15 kb fragment
Lane 2 : 20 kb fragment
Lane 3 : 25 kb fragment
Lane 4 : 30 kb fragment

MSDS
• AccuPower® ProFi Taq PCR PreMix

Brochure
• AccuPower® PreMix series - 2010 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

AccuPower® Gold Multiplex PCR PreMix can amplify up to 20 target genes in a single tube. AccuPower® Gold Multiplex PCR PreMix contains Bioneer’s unique enzyme-mediated HotStart technology with Pyrophosphatase (PPase) and Pyrophosphate (PPi) for efficient suppression of non-specific products and enhanced amplification specificity.
AccuPower® Gold Multiplex PCR PreMix can be used for a variety of applications including genotyping assays or molecular diagnostics, and can also be used for cDNA-based semi-quantitative assays.

 

Features and Benefits

• Flexibility: Up to 20 different target genes from human genomic DNA can be amplified in a single tube
• Specificity: Pyrophosphate (PPi) has high affinity for Mg2+. By adding PPi to the reaction mixture, the Mg2+ ions necessary for normal PCR are bound, preventing DNA polymerase activity. This PPi-Mg2+ binding prevents non-specific before PCR (zero-cycle) product formation. Upon thermal cycling, the pyrophosphatase (PPase) that is also added to the mixture is activated (>70°C) and hydrolyzes the PPi to 2 phosphate groups and facilitates the release of Mg2+, which is then available for DNA polymerase to use and resume normal activity.
• Easy-to-use: All reaction components required for PCR, including thermostable DNA polymerase and dNTPs are contained within each tube and in a lyophilized “PreMix” form. The user needs only to add template DNA, primers and water to perform up to 20-plex PCR. Materials necessary for loading agarose gels for electrophoresis are also added in the reaction, negating the need to add loading dye after PCR is completed.
• Reproducibility: Each batch is produced under strict quality controls. Errors that may occur during mass production are eliminated during the individual packaging process. Bioneer’s current batch processing system allows for the production of more accurate and reproducible end-product. Additionally, the streamlining of setup using a lyophilized premix enhances reproducibility by minimizing setup variables.

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: Yes
Fragment Size: Up to 1 kb


Application

Result


Figure 1. High specificity of AccuPower® Gold Multiplex PCR PreMix
Each lane from left to right represensts the progressive number of primer sets (1 – 20) included in single AccuPower® Gold Multiplex PCR PreMix.

Figure 2. Single PCR and Multiplex PCR using AccuPower® Gold Multiplex PCR PreMix
Each lane from left to right indicates the single and multiplex PCR products using AccuPower® Gold Multiplex PCR PreMix.
a) 10-plex PCR, b) 20-plex PCR

Figure 3. Comparison of amplification quality between AccuPower® Gold Multiplex PCR PreMix and competitor Multiplex PCR kits.
6-plex (a), 10-plex (b), 20-plex (c) primers were added into AccuPower® Gold Multiplex PCR PreMix and competitor master mixtures. A series of human genomic DNA diluents were tested (Lane 1: 100 ng, Lane 2: 10 ng, Lane 3: 1 ng).
 

Figure 4. Comparison of amplification quality using Labchip® between AccuPower® Gold Multiplex PCR PreMix and competitor Multiplex PCR kits.
a) Virtual Gel Image. Gel image illustrates data reproducibility of the LabChip 90 system.
b) Overlay of expression levels using Bioneer’s AccuPower® Gold Multiplex PCR PreMix and competitor multiplex PCR kits. The electropherogram displays the data between 10 PCR product yields using a 10-plex primer set to illustrate amplification efficiency.
c) The graph shows the total concentration of PCR products between AccuPower® Gold Multiplex PCR PreMix and other supplier’s master mixtures.            

MSDS
• AccuPower® Gold Multiplex PCR PreMix

Brochure
• AccuPower® Gold Multiplex PCR PreMix - 2011 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

Features and Benefits

• Fexibility: Up to 20 different target genes from human genomic DNA can be amplified in a single tube
• Easy-to-use:  All reaction components required for PCR, including thermostable DNA polymerase and dNTPs are contained within each tube and in a lyophilized “PreMix” form. The user needs only to add template DNA, primers and water to perform up to 20-plex PCR. Materials necessary for agarose gel electrophoresis are also added in the reaction, negating the need to prepare the samples for electrophoresis after PCR is completed.
• Reproducibility: Each batch is produced under strict quality controls. Errors that commonly occur during mass production are eliminated during the individual packaging process. Bioneer’s current batch processing system allows for the production of more accurate and reproducible end-product yield.

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: Yes
Fragment Size: ~1 kb

Application
 

Figure 1. Single PCR and Multiplex PCR using AccuPower® Multiplex PCR PreMix

Each lane from left to right indicates the single and Multiplex PCR product using AccuPower® Multiplex PCR PreMix.
a) 10plex Multiplex PCR, b) 20plex Multiplex PCR
M; 25/100 bp Mixed DNA Ladder (cat. No. D-1020)


Figure 2. Comparison of amplification quality between AccuPower® Multiplex PCR PreMix and other supplier’s Multiplex PCR kit.

6-plex primers were added into AccuPower® Multiplex PCR PreMix and other supplier’s Multiplex PCR kit. A series of Human genomic DNA diluents were tested. ( Lane 1. Human genomic DNA 100ng, Lane 2. Human genomic DNA 10ng, Lane 3. Human genomic DNA 1ng ). All data were obtained using Mygene 96 machine (Applied Bioneer co.)
Supplier Q : Q company Multiplex PCR kit, Supplier S : S company Multiplex PCR kit, Supplier I : I company Taq polymerase for Mutiplex PCR (0.5U, added 2mM MgCl₂) Rxn. Condition : 95°C for 10min, followed by 35cycles of 30sec at 95°C, 30sec at 60°C, 60sec at 72°C M; 25/100 bp Mixed DNA Ladder (cat. No. D-1020)

Figure 3. Comparison of amplification quality between AccuPower® Multiplex PCR PreMix and other supplier’s Multiplex PCR kit.

10-plex primers were added into AccuPower® Multiplex PCR PreMix and other supplier’s Multiplex PCR kit. A series of Human genomic DNA diluents were tested. ( Lane 1. Human genomic DNA 100ng, Lane 2. Human genomic DNA 10ng, Lane 3. Human genomic DNA 1ng ). All data were obtained using Mygene 96 machine (Applied Bioneer co.)
Supplier Q : Q company Multiplex PCR kit, Supplier S : S company Multiplex PCR kit, Supplier I : I company Taq polymerase for Mutiplex PCR (0.5U, added 2mM MgCl₂)Rxn. Condition : 95°C for 10min, followed by 35cycles of 30sec at 95°C, 30sec at 65°C, 60sec at 72°C M; 25/100 bp Mixed DNA Ladder (cat. No. D-1020)

Figure 4. Comparison of amplification quality using Lapchip® between AccuPower® Multiplex PCR PreMix and other supplier’s master mixture.

a) Virtual Gel Image. Gel image illustrates data reproducibility of the LabChip 90 system.
b) Overlay of expression level using Bioneer’s multiplex PCR PreMix and other company’s multiplex PCR kit. The electropherogram displays the data between 10 PCR products yield using 10-plex primer sets to illustrate the Amplification efficiency.
c) The graph shows the total concentration of PCR products between AccuPower® Multiplex PCR PreMix and other supplier’s master mixture.


Figure 5. Comparison of amplification quality between AccuPower® Multiplex PCR PreMix and other supplier’s Multiplex PCR kit.

20-plex primers were added into AccuPower® Multiplex PCR PreMix and other supplier’s Multiplex PCR kit. A series of Human genomic DNA diluents were tested. (100ng ~ 1ng). All data were obtained using Mygene 96 machine (Applied Bioneer co.)
Supplier Q : Q company Multiplex PCR kit, Supplier S : S company Multiplex PCR kit, Supplier I : I company Taq polymerase for Mutiplex PCR (0.5U, added 2mM MgCl₂)
Rxn. Condition : 95°C for 10min, followed by 35cycles of 30sec at 95°C, 30sec at 57°C, 60sec at 72°C M; 25/100 bp Mixed DNA Ladder (cat. No. D-1020) Trademark: 'AccuPower®' is a registered trademark of Bioneer Corporation.

MSDS
• AccuPower® Multiplex PCR PreMix

Brochure
• Amplification New Product-2011 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

Features and Benefits

Thermostable Activity:
Original M-MLV RTase has maximum activity at relatively low temperatures (42 °C), causing several problems in reverse transcription of complex secondary structure RNA molecules. To remedy this issue, Bioneer has developed a RTase active at high temperatures (above 50°C). RocketScript™ has thermostable activity (42~70°C), allowing efficient cDNA synthesis from complex secondary structure RNA and give the user freedom to optimize the reverse transcription reaction based on temperature.

Ease-of-Use:
- The product contains our thermostable RTase RocketScript™ , RNase inhibitors and all other components necessary for
a successful RT reaction in a single tube. Just add RNA, primers and PCR-grade water to perform the RT reaction.
-Components necessary for agarose gel electrophoresis are also contained within the product including tracking dye and
a density-increasing reagent for added convenience.
-T vector cloning is possible without any additional reactions.

Reproducibility:
Each product batch is produced under strict quality control processes for accurate and reproducible end product manufacturing.

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: Yes
Fragment Size: 6 kb

Application

- First-strand synthesis of cDNA from RNA molecules (RT)
- RT-PCR
- Library construction
- Gene-expression analysis

Experimental Data


Figure 1. Performance comparison between AccuPower® RocketScript™ RT-PCR PreMix and competitor RT-PCR kits

Complex secondary structure RNA species amplification by RocketScript™ is outstanding compared to the leading competitor’s reverse transcriptase. RT reactions were performed according to each manufacturer’s recommendations.
Lanes 1-4 are 10 ng, 1 ng, 0.1 ng and 0.01 ng of total RNA from HeLa cells, respectively

Figure 2. AccuPower® RocketScript™ RT-PCR PreMix shows enhanced performance compared to competitors

One-Step RT-PCR reactions were performed with 100ng total RNA from HeLa cells using regents and conditions specified in each manufacturer’s protocol
Lane 1: 2.5kb Lane 2: 3kb Lane 3: 4.5kb Lane 4: 5.2kb

MSDS
• AccuPower® RocketScript™ RT-PCR PreMix

Brochure
• AccuPower® RocketScript™ RT-PCR PreMix - 2011 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

For cDNA amplification with Cyclic RT, please see our AccuPower® CycleScript RT PreMix.
 

Features and Benefits

•  Convenient lyophilized RT: Easy to use, simply add your purified RNA
•  High yield of cDNA: For genes up to 9 kb within 10 minutes
•  Stable for 2 years at -20°C: Long life
•  RNase, DNase and Proteinase-free: Ensures the integrity of your samples

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: No
Fragment size: 9 kb

Application

cDNA synthesis



Figure 1. Reliability and reproducibility test with AccuPower® RT PreMix. Each template showcases amplified target genes.

Lane M: 100 bp DNA Ladder (D-1030)
Lane 1-4: Reliability test of each lot with AccuPower® RT PreMix

MSDS
• AccuPower® PCR Master Mixes

Brochure
• AccuPower® PCR Master Mixes - 2010 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

AccuPower® RocketScript™ RT PreMix contains BIoneer’s exclusive M-MLV based thermostable reverse transcriptase (RTase), RocketScript™. Native M-MLV RTase has maximum activity at relatively low temperatures (42°C), causing several problems in reverse transcription of RNA molecules with complex secondary structure.

RocketScript™ has thermostable activity (42°C~70°C), allowing efficient cDNA synthesis from virtually any RNA. The lyophilized PreMix contains all components necessary for a successful reverse transcription reaction, including RTase, RNase inhibitor and buffer components. Just add template RNA, primers and water, and the RT reaction is ready to go.

Features and Benefits

• Thermostable Activity: RocketScript™ is able to perform reverse transcription reactions throughout a wide range of temperatures from 42°C to 70°C.

• Enhanced Performance: RocketScript™ has enhanced performance to handle both high and low input RNA concentrations as well as short and long RT target sizes.

• Easy-to-use: The product contains the enzyme itself, plus RNase inhibitors and all other components necessary for the best reverse transcription results in the tube.

• Reproducibility: Bioneer's strict quality controlled production system ensures that your results will be reproducible experiment after experiment.

• Convenient: Just add template and primers and start your reaction. dNTPs, buffer and enzyme are provided.

• Stability: Stable at room temperature for a month one year at 4°C and for 2 years in a -20°C freezer

• RNase, DNase and Proteinase-free: Ensures the integrity of your samples.

• Broad range of working temperatures: For RNAs that are G:C rich or significant secondary structure

• Stable for 2 years at -20°C: Long shelf life

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: No
Fragment Size: Up to 10 kb

Application

- First-strand synthesis of cDNA from RNA molecules (RT)
- RT-PCR
- Random priming reaction
- Library construction
- Probe labeling
- mRNA 5end Mapping by Primer Extension Analysis
- Real time PCR

Experimental data

Figure 1. Sensitivity comparison between AccuPower® RocketScript™ RT PreMix and M-MLV RTase
Sensitivity results of AccuPower® RocketScript™ RT PreMix using GAPDH compared with conventional Reverse transcriptases.
Each 100 ng – 10 fg of total RNA used for RT and the same amount of RT products used for electrophoresis.
Lane 1: 10 fg Human total RNA from HeLa cell Lane 2: 100 fg Human total RNA from HeLa cell
Lane 3: 1 pg Human total RNA from HeLa cell Lane 4: 10 pg Human total RNA from HeLa cell
Lane 5: 100 pg Human total RNA from HeLa cell Lane 6: 1 ng Human total RNA from HeLa cell
Lane 7: 10 ng Human total RNA from HeLa cell Lane 8: 100 ng Human total RNA from HeLa cell


Figure 2. Comparison of amplification efficiency between AccuPower® RocketScript™ RT PreMix and competitors M-MLV RTase.  
    RocketScript™ is able to handle a wide range of sample concentrations and transcript lengths so your downstream applications are minimally effected by the reverse transcription step.
Lanes 1-3: 1,000 ng, 100 ng and 10 ng of total RNA from HeLa cells, respectively.

Figure 3. Sensitivity comparison between AccuPower® RocketScript™ RT PreMix and competitor RTases using Real Time PCR
Reverse transcription conditions: conventional 1 hr incubation at 60°C, deactivation at 95°C for 5 min All cDNAs were amplified with AccuPower® DualStar™ qPCR PreMix (K-6110) from Bioneer.

MSDS
• AccuPower® RocketScript RT PreMix

Brochure
• AccuPower® RocketScript RT PreMix - 2011 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

AccuPower® RocketScript™ Cycle RT PreMix contains Bioneer’s exclusive M-MLV based thermostable reverse transcriptase
(RTase), RocketScript™. Conditions are optimized for Bioneer’s Cyclic Temperature Reverse Transcription (CTRT; patent pending) in a PreMix form.

Native M-MLV RTase has maximum activity at relatively low temperatures (42°C), causing several problems in reverse
transcription of RNA molecules with complex secondary structure.
AccuPower® RocketScript™ Cycle RT PreMix has thermostable activity across a wide temperature range, (42°C – 70°C),
allowing efficient cDNA synthesis from virtually any RNA.
The lyophilized PreMix contains all components necessary for a successful CTRT reaction, including RTase, RNase inhibitor and buffer components. Just add template RNA, primers and water, and the RT reaction is ready to go.

Schematic representation of the 5’UTR of a gene, with complex secondary structure, at three different temperatures.
Note that RocketScript™ shows full activity at 70°C allowing it to synthesize the complete gene sequence where M-MLV and other Reverse Transcriptase's fail.
 

Features and Benefits 

• Thermostable Activity: RocketScript™ is able to perform reverse transcription reactions throughout a wide range of temperatures from 42°C to 70°C.

• Enhanced Performance: RocketScript™ has enhanced performance to handle both high and low input RNA concentrations as well as short and long RT target sizes.

• Easy-to-use: The product contains the enzyme itself, plus RNase inhibitors and all other components necessary for the best reverse transcription results in the tube.

• Reproducibility: Bioneer's strict quality controlled production system ensures that your results will be reproducible experiment after experiment.

• Convenient: Just add template and primers and start your reaction. dNTPs, buffer and enzyme are provided.

• Stability: Stable at room temperature for a month one year at 4°C and for 2 years in a -20°C freezer

• RNase, DNase and Proteinase-free: Ensures the integrity of your samples.

• Broad range of working temperatures: For RNAs that are G:C rich or significant secondary structure

• Stable for 2 years at -20°C: Long shelf life

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: No
Fragment Size: Up to 10 kb

Application

- First-strand synthesis of cDNA from RNA molecules (Reverse Transcription)
- RT-PCR
- Random priming reaction
- Library construction
- Probe labeling
- mRNA 5' end mapping by primer extension analysis
- Real time PCR

Experimental data

Figure 1. Comparison of amplification efficiency between AccuPower® RocketScript™ Cycle RT PreMix and competitors’ RTases
(a) Sensitivity test
Primer set: Human TFRC set
Lane M: 1 kb DNA Ladder
Lane1: 100 ng Human total RNA from HeLa cell
Lane 2: 10 ng Human total RNA from HeLa cell
Lane3: 1 ng Human total RNA from HeLa cell
Lane4: 100 pg Human total RNA from HeLa cell
RT reaction condition is performed according to each manufacturer’s recommendations

(b) Full-Length cDNA synthesis test
RT reactions were performed according to each manufacturer’s recommendation. All cDNAs were amplified with AccuPower® Hotstart PCR Premix (K-5050) from Bioneer
Note: Competitor products show inhibition at high input concentrations of total RNA
Lane 1: 1 ug Human total RNA from HeLa cell
Lane 2: 100 ng Human total RNA from HeLa cell
Lane 3: 10 ng Human total RNA from HeLa cell


Figure 2. Complex RNA amplification results of RocketScript™ Cycle RT PreMix
Each target gene was amplified after performing reverse transcription with AccuPower® RocketScript™ Cycle RT PreMix.
Reverse transcription conditions: Conventional 1 hr incubation at 42°C, 50°C, or 60°C, deactivation at 95°C for 5 min
A: M-MLV Reverse Transcriptase
B: AccuPower® RocketScript™ Cycle RT PreMix with Oligo (dT)20
Lane 1: 100 ng Human total RNA from HeLa cell
Lane 2: 10 ng Human total RNA from HeLa cell
 

FTRT (Fixed Temperature Reverse Transcription) conditions:

CTRT (Cyclic Temperature Reverse Transcription) conditions:

Primer set: Human GAPDH and myc set
Lane M: 1 kb DNA Ladder
Lane1: 10 ng Human Total RNA from HeLa Cell
Lane 2: 1 ng Human Total RNA from HeLa Cell
Lane 3: 100 pg Human Total RNA from HeLa Cell
Lane 4: 10 pg Human Total RNA from HeLa Cell


Figure 5. Comparison of amplification results between Bioneer RocketScript™ Cycle RT PreMix and competitor RTases
All cDNAs were amplified with AccuPower® Hotstart PCR Premix (K-5050) from Bioneer
Primer set: human Myc498 bp set
Lane M: 1 kb DNA Ladder
Lane1: 10 ng Human total RNA from HeLa Cell
Lane 2: 1 ng Human total RNA from HeLa Cell
Lane 3: 100 pg Human total RNA from HeLa Cell
Lane 4: 10 pg Human total RNA from HeLa Cell

MSDS
• AccuPower® RocketScript Cycle RT PreMix

Brochure
• AccuPower® RocketScript Cycle RT PreMix - 2011 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

AccuPower® CycleScript RT was developed for both a conventional reverse transcription PCR at a fixed temperature at 42°C as well as Cyclic Reverse Transcription that is carried out like a PCR (please see the following comparison table). Bioneer’s Cyclic RT is an innovative technology to synthesize more homogeneous cDNA in less time compared to the conventional reverse transcription.
 

Conventional Reverse Transcription

Cyclic Reverse Transcription

Features and Benefits

•  Convenient lyophilized RT premix: Easy to use, simply add your purifies RNA
•  Broad range of working temperatures: For RNAs that are G:C rich or significant secondary structure
•  Novel Cyclic Reverse Transcription: Ensures that you isolate even the rarest transcripts
•  High yield of cDNA:  For genes up to 9 kb within 10 minutes
•  Stable for 2 years at -20°C:  Long life
•  RNase, DNase and Proteinase-free: Ensures the integrity of your samples

3' to 5' exonuclease: No
3’ – A Overhang: No
Fragment size: 9 kb

Application

cDNA synthesis



Figure 1. Reaction condition comparison of Cyclic Reverse Transcription versus conventional reverse transcription PCR. 800 ng of HeLa cell total RNA was reverse transcribed using different reaction conditions and then 800 bp, 1.5 kb, 2.0 kb, and 2.6 kb of human transferrin receptor gene were amplified using AccuPower® PCR PreMix.

M: 1 kb DNA Ladder (D-1040)
Lane 1 - 4: Conventional RT with dT₂₀ at 42°C
Lane 5 – 8: Conventional RT with dN₁₂ at 42°C
Lane 9 - 12: Conventional RT with dT₂₀ at 55°C
Lane 13 - 16: Conventional RT with dN₁₂ at 55°C
Lane 17 - 20: Cyclic RT with dT₂₀ by 55°C
Lane 21 - 24: Cyclic RT with dN₁₂ at 55°C




Figure 2. Coxsackie virus A9 (AJ295200) 5’-UTR region amplification with different reverse transcription products. Coxsackie virus RNA was reverse transcribed with different reverse transcription products and then amplified with PCR with AccuPower® PCR PreMix. All materials were used in accordance with manufacturer’s instructions.

M: 100 bp DNA Ladder (D-1030)
Lane 1: Reverse transcription with AccuPower® RT PreMix and oligo dT primer
Lane 2: Reverse transcription with AccuPower® RT PreMix and random primer
Lane 3 – 4, 7 – 8, 11 – 12: Reverse transcription with AccuPower® CycleScript RT PreMix and dT₂₀
Lane 5 – 6, 9 – 10, 13 – 14: Reverse transcription with AccuPower® CycleScript RT PreMix and dN₁₂
Lane 15 – 16: Reverse transcription with RT product from company C and oligo dT primer
Lane 17 – 18: Reverse transcription with RT product from company C and random primer
Lane 19 – 20: Reverse transcription with RT product from company I and oligo dT primer
Lane 21 – 22: Reverse transcription with RT product from company I and random primer




Figure 3. Long transcript production comparison. Reverse transcription was carried out with various reverse transcription products prior to PCR with AccuPower® PCR PreMix.

M: 100 bp DNA Ladder (D-1030)
Bioneer: Bioneer / 1 hour incubation at 55°C
C: Company C / 1 hour incubation at 42°C
I: Company I / 1 hour incubation at 45°C
Lane 1, 3, 5: Product reverse transcribed with oligo dT primer
Lane 2, 4, 6: Product reverse transcribed with random primer
 

Figure 4. Transferrin receptor gene amplification with various reverse transcription products. Reverse transcription was carried out with four different reverse transcription products according to manufacturer’s instructions and then cDNA was amplified with AccuPower® PCR PreMix.

M: 1 kb DNA Ladder (D-1040)
Lane 1 – 5: Reverse transcribed with AccuPower® CycleScript RT PreMix (dT₂₀) / 1 hour at 55°C
Lane 6 – 10: Reverse transcribed with AccuPower® CycleScript RT PreMix (dN₁₂) / 1 hour at 55°C
Lane 11 – 15: Reverse transcribed with Company C product and oligo dT primer / 1 hour at 42°C
Lane 16 – 20: Reverse transcribed with Company C product and random primer / 1 hour at 42°C
Lane 21 – 25: Reverse transcribed with Company I product and dT primer / 1 hour at 45°C
Lane 26 – 30: Reverse transcribed with Company I product and random primer / 1 hour at 45°C
 

Figure 5. Reaction temperatures and time comparison. Data A and B demonstrate high thermal stability of AccuPower® CycleScript RT PreMix.

A: Reverse transcription carried out with AccuPower® CycleScript RT PreMix and dT₂₀
B: Reverse transcription carried out with AccuPower® CycleScript RT PreMix and dN₁₂
Conventional RT #1: 1 hour incubation at 42°C
Cyclic RT #1: Repeat 12 times of 2 minutes at 37°C and 3 minutes at 50°C
Conventional RT #2: 1 hour incubation at 55°C
Cyclic RT #2: Repeat 12 times of 1 minutes at 37°C, 3 minutes at 47°C, and 3 minutes at 55°C
M: 100 bp DNA Ladder (D-1030)
Lane 1, 5, 9, 13, 17, 21, 25, 29: 100 ng of HeLa cell total RNA used for reaction
Lane 2, 6, 10, 14, 18, 22, 26, 20: 10 ng of HeLa cell total RNA used for reaction
Lane 3, 7, 11, 15, 19, 23, 27, 31: 1 ng of HeLa cell total RNA used for reaction
Lane 4, 8, 12, 16, 20, 24, 28, 32: 100 pg of HeLa cell total RNA used for reaction
 

Figure 6. Data C and D demonstrate superior productivity of AccuPower® CycleScript RT PreMix..

C: Reverse transcription carried out with AccuPower® CycleScript RT PreMix and dT₂₀
D: Reverse transcription carried out with AccuPower® CycleScript RT PreMix and dN₁₂
10 min: 2 times of 2 minutes at 37°C and 3 minutes at 50°C
20 min: 4 times of 2 minutes at 37°C and 3 minutes at 50°C
60 min: 12 times of 2 minutes at 37°C and 3 minutes at 50°C
M: 100 bp DNA Ladder (D-1030)
Lane 1, 5, 9, 13, 17, 21: 100 ng of HeLa cell total RNA used for reaction
Lane 2, 6, 10, 14, 18, 22: 10 ng of HeLa cell total RNA used for reaction
Lane 3, 7, 11, 15, 19, 2: 1 ng of HeLa cell total RNA used for reaction
Lane 4, 8, 12, 16, 20, 24, 28, 32: 100 pg of HeLa cell total RNA used for reaction

• AccuPower® CycleScript RT PreMix (dN12)

• AccuPower® CycleScript RT PreMix (dN20)

MSDS
• AccuPower® PCR Master Mixes

Brochure
• AccuPower® PCR Master Mixes - 2010 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

Features and Benefits

• Convenient lyophilized RT: Easy to use, simply add your purified RNA
•  High yield of cDNA: For genes up to 6 kb
•  Stable for 2 years at -20°C: Long life
•  RNase, DNase and Proteinase-free: Ensures the integrity of your samples
   

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: No
Fragment size: 6 kb

Application

cDNA synthesis followed by PCR

Lane 1: DNA Ladder (BM corp.)
Lane 2: Negative control
Lane 3, 4: HCV positive serum
Lane 5: HCV negative serum
 

Lane1: 10⁹ copy Lane2: 10⁸ copy
Lane3: 10⁷ copy Lane4: 10⁶ copy
Lane5: 10⁵ copy Lane6: 10⁴ copy
Lane7: 10³ copy Lane8: NTC
 

Lane 1: 10 ng Human total RNA from HeLa cell
Lane 2: 1 ng Human total RNA from HeLa cell
Lane 3: 100 pg Human total RNA from HeLa cell
Lane 4: 10 pg Human total RNA from HeLa cell

MSDS
• AccuPower® PCR Master Mixes

Brochure
• AccuPower® PCR Master Mixes - 2010 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

The polymerase chain reaction (PCR) can amplify a single molecule over a billionfold. Thus, even minuscule amounts of a contaminant can be amplified and lead to a false positive result. Such contaminants are often products from previous PCR amplifications (carry-over contamination). Therefore, researchers have developed methods to avoid such contamination AccuPower® Hotstart PCR PreMix(with UDG) is a ready-to-use master mix for high-specificity DNA amplification, con-taining UDG to prevent carryover contamination or crossover contamination The master mix combines the Enzyme - mediated HotStart technology of HotStart DNA polymerase with integrated UDG carryover prevention technology to provide optimal performance with a variety of PCR detection technologies
 

Features and Benefits

• Prevention of carryover contamination

• Specificity

•  Easy-to-use

•  Reproducibility

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: Yes
Fragment Size: ~ 12 kb

Application

Genome template’s PCR
Low-copy target's PCR
Multiple primer pair's PCR
cDNA templates PCR
Molecular Diagnosis


Figure 1. Comparison of specificity between Accupower® HotStart PCR PreMix and HotStart PCR PreMix(with UDG).
Specificity test was operated using 7 pair of primers targeting P53 gene. Reaction mixture was incubated at 37°C for 2min followed by 95°C for 5min, 30 cycles of 20sec at 95°C, 40sec at 55°C, 1min at 72°C. Accupower® HotStart PCR PreMix(with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP. The amount of DNA(human) used to test is 10ng and size of PCR amplicon size each lane is represented below.
Lane 1: 139bp Lane 2: 211bp
Lane 3: 447bp Lane 4: 618bp
Lane 5: 1082bp Lane 6: 1296bp
Lane 7: 1561bp
M: 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)


Figure 2. Comparison of sensitivity between Accupower® HotStart PCR PreMix and HotStart PCR PreMix(with UDG).
Sensitivity test was operated using serial diluted human genomic DNA. Reaction mixture was incubated at 37°C for 2min followed by, 95°C for 5min, 30 cycles of 20sec at 95°C, 20sec at 55°C, 30sec at 72°C. Accupower® HotStart PCR PreMix(with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP. The amount of DNA used to test is represented below.
Lane 1: 10ng of Human genomic DNA Lane 2: 1ng of Human genomic DNA
Lane 3: 100pg of Human genomic DNA Lane 4: 10pg of Human genomic DNA
M: 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)


Figure 3. Comparison of amplification quality using PCR products(not including uracil base) between Accupower® HotStart PCR PreMix and Accupower® HotStart PCR PreMix(with UDG).
Amplification quality test was operated using serial diluted PCR products not including uracil base. Accupower® HotStart PCR PreMix was also tested for negative control. Reaction mixture was incubated at 37/7°C for 2min followed by 95°C for 5min, 30 cycles of 20sec at 95°C, 20sec at 55°C, 30sec at 72°C. Accupower® HotStart PCR PreMix(with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP. The copy number of PCR products used to test are represented below.
Lane 1: 10¹¹copy Lane 2: 10¹⁰copy Lane 3: 10⁹copy
Lane 4: 10⁸copy Lane 5: 10⁷copy Lane 6: 10⁶copy
Lane 7: 10⁵copy Lane 8: 10⁴copy Lane N: No template control
M: 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)

Figure 4. Efficiency of uracil DNA glycosylase using PCR product(including uracil base).
Efficiency test of uracil DNA glycosylase was operated using serial diluted PCR products including uracil base. Accupower® HotStart PCR PreMix was also tested for negative control. Reaction mixture was incubated at 37°C for 2min followed by 95°C for 5min, 30 cycles of 20sec at 95°C, 20sec at 55°C, 30sec at 72°C. Accupower® HotStart PCR PreMix(with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP.
The copy number of PCR products used to test are represented below.
Lane 1: 10¹¹copy Lane 2: 10¹⁰copy Lane 3: 10⁹copy
Lane 4: 10⁸copy Lane 5: 10⁷copy Lane 6: 10⁶copy
Lane 7: 10⁵copy Lane 8: 10⁴copy Lane N: No template control
M: 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)

• AccuPower® Hotstart PCR PreMix(with UDG)

MSDS
• AccuPower® Hotstart PCR PreMix(with UDG)

    
Brochure
• AccuPower® PreMix series - 2010 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

Features and Benefits

• Specificity

• Stability

Stable at room temperature for a month and for 2 years in a -20°C freezer
 

• Ease-of-Use

All reaction components required for PCR, including thermostable DNA polymerase and dNTPs are contained within each tube and in a lyophilized "PreMix" form. The user needs only to add template DNA, primers and water. Materials necessary for loading agarose gels for electrophoresis are also added in the reaction, negating the need to add loading dye after PCR is completed.
 

• Reproducibility

5' to 3' exonuclease: Yes
3' to 5' exonuclease: No
3' – A Overhang: Yes
PCR product size: ~ 5 kb(Human)

Application

- High specificity PCR
- High sensitivity PCR
- Low-copy target PCR
- Multiple primer pairs PCR
- cDNA template PCR
- T-A cloning

Experimental data

Figure 1. Comparison of PCR amplification specificity between AccuPower® PyroHotStart Taq PCR PreMix from Bioneer and other suppliers' Hot start PCR master mix.

PCR reactions were performed according to each supplier's protocol. The PrP gene was amplified from human genomic DNA with two different primer sets, separately. This data shows that AccuPower® PyroHotStart Taq PCR PreMix has higher amplification efficiency and specificity than other suppliers' Hot start PCR master mix.
Lane M: 100 bp DNA Ladder(Bioneer, Cat. No.D-1030)
Lane 1: 100 ng DNA, PrP primer set (500 bp)
Lane 2: 10 ng DNA, PrP primer set (500 bp)
Lane 3: 100 ng DNA, PrP primer set (705 bp)
Lane 4: 10 ng DNA, PrP primer set (705 bp)

Figure 2. Comparison of PCR amplification specificity between AccuPower® PyroHotStart Taq PCR PreMix from Bioneer and other suppliers' Hot start PCR master mix.

The ApoE gene was amplifed from 100 ng of human genomic DNA (The PCR product size is 268bp). This data shows that AccuPower® PyroHotStart Taq PCR PreMix has higher amplificition efficency and specificity than other suppliers' Hot start PCR master mix.
Lane M: 100 bp DNA Ladder (Bioneer, Cat. No. D-1030)
Lane 1: AccuPower® PyroHotStart Taq PCR PreMix
Lane 2: Supplier I Hotstart Taq PCR preMix
Lane 3: Supplier S Hotstart Taq PCR masterMix
Lane 4: Supplier T Hotstart Taq PCR masterMix
Lane 5: Supplier Q Hotstart Taq PCR masterMix

Figure 3. AccuPower® PyroHotStart Taq PCR PreMix has high amplification efficiency and specificity.

Specificity test was performed using 7 different sets of primers targeting the P53 gene. 10 ng of human genomic DNA was used for each PCR reaction. The cycling conditions were 95°C for 5min, 30 cycles of 95°C for 20 sec, 55°C for 40 sec, and 72°C for 1 min, and 72°C for 5 min for final extension.
Lane M: 100bp DNA Ladder (Bioneer, Cat. No. D-1030)
Lane 1: P75/73 primer set (139 bp)
Lane 2: P55/53 primer set (211 bp)
Lane 3: P55/63 primer set (447 bp)
Lane 4: P75/83 primer set (618 bp)
Lane 5: P55/73 primer set (1082 bp)
Lane 6: P65/83 primer set (1296 bp)
Lane 7: P55/83 primer set (1561 bp)

Figure 4. Comparison of PCR amplification between AccuPower® PyroHotStart Taq PCR PreMix from Bioneer and other suppliers' Hot start PCR master mix.

Sensitivity test was performed by amplifying the IRGC gene from a serial dilution of human genomic DNA. This data shows that AccuPower® PyroHotStart Taq PCR PreMix has higher amplification efficiency than other suppliers' Hot start PCR master mix.
Lane M: 100 bp DNA Ladder (Bioneer, Cat. No. D-1030)
Lane 1: 10 ng human genomic DNA
Lane 2: 1 ng human genomic DNA
Lane 3: 100 pg human genomic DNA
Lane 4: 10 pg human genomic DNA

Figure 5. Comparison of cDNA template amplification between AccuPower® PyroHotStart Taq PCR PreMix and other suppliers' Hot start PCR master mix.

cDNA synthesized from 10-fold serial-diluted human total RNA from 10 ng to 10 pg using AccuPower® RocketScript™ Cycle RT PreMix (Bioneer, Cat. No. K-2201) was used as a template for PCR amplification. This data shows that AccuPower® PyroHotStart Taq PCR PreMix has higher amplification efficiency than other suppliers' Hot start PCR master mix.
Lane M: 100 bp DNA Ladder (Bioneer, Cat. No. D-1030)
Lane 1: 10 ng human total cDNA
Lane 2: 1 ng human total cDNA
Lane 3: 100 pg human total cDNA
Lane 4: 10 pg human total cDNA

MSDS
• AccuPower® PyroHotstart Taq PCR PreMix

Brochure
• AccuPower® PreMix series - 2010 Brochure

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

Features and Benefits

• User Convenience: Just add probe and primer for target gene.
• Extended Stability: In a freezer, the activity of this product is stable for 2 years.
• Reproducibility: Margin of error was minimized and reproducible results can be obtained, because no extra mixing step is requied.
• High Specificity: A Non-specific reaction by Enzyme-mediated Hotstart Technology was dramatically eliminated(patent 292883 & 10-2007-109055)
• Equipment Compatibility: Optimized results on various Real-Time PCR instruments can be obtained.
• Universality of target gene: Significant results can be obtained regardless of kinds of target genes.

5' to 3' exonuclease: Yes
3' to 5' exonuclease: No
3’ – A Overhang: Yes
Fragment size: ~1 kb

Application

Real-Time quantification of DNA and cDNA targets, gene expression profiling, microbial & viral pathogen detection, evaluation of probe-based real-time PCR system


Figure 1. Real-Time PCR Data of AccuPower® Dualstar™ qPCR PreMix

AccuPower® Dualstar™ qPCR PreMix provides at least 7 orders of magnitude in dynamic range (10⁷ ~ 10¹ copies/reaction).
A : Amplification curve. West Nile Virus (WNV) primers and TaqMan-based probe were added into Dualstar qPCR Premix.
A series of WNV positive control diluents were tested.
B : Standard curve. All data were obtained using Exicycler™ 96 Real-Time Quantitative Thermal Block (Bioneer Co.).


Figure 2. Data using various kinds of Real-Time PCR Instruments (ABI 7500 Fast, ABI 7500, Opticon Real-Time PCR instruments)

AccuPower® Dualstar™ qPCR PreMix provides at least 7 orders of magnitude in dynamic range (10⁷ ~ 10¹ copies/reaction).
A : Amplification curve. West Nile Virus (WNV) primers and TaqMan-based probe were added into Dualstar qPCR Premix.
A series of WNV positive control diluents were tested.
B : Standard curve. All data were obtained using Exicycler™ 96 Real-Time Quantitative Thermal Block (Bioneer Co.).

Notice to Purchaser

FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.

The lyophilized composition of this product is covered by US Patent Nos. 5861251 and 6153412 of Bioneer Corporation. Certain applications of this product are covered by pending or issued patents in certain countries. Because purchase of this product does not include a license to perform any patented application, users of this product may be required to obtain a patent license depending upon the particular application and country in which the product is used. Products may be covered by pending or issued patents. Please contact us for more information

Trademark: 'AccuPower®' is a registered trademark of Bioneer Corporation. 'DualStar™' is a trademark of Bioneer Corporation.

* Some of the products may not be available in some countries, For details, please contact your local distributor.

MSDS
• AccuPower® Dualstar™ qPCR PreMix

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

The Real-Time PCR Reagents for the SYBR® Green-Based Detection



The Greenstar™ qPCR PreMix provides significant reduction of nonspecific reaction, high sensitivity, extended stability and great universality. Just additions of primers of your interesting target gene into Greenstar™ qPCR PreMix provide always reproducible results with convenience of use.

Features and Benefits

• High Specificity: A Non-specific reaction by Enzyme-mediated Hotstart Technology was dramatically eliminated(patent 292883 & 10-2007-109055)
• User Convenience:  Just add primer for target gene.
• Extended Stability: In a freezer, the activity of this product is stable for 2 years.
• Reproducibility: Margin of error was minimized and reproducible results can be obtained, because no extra mixing step is requied.
• Equipment Compatibility: Optimized results on various Real-Time PCR instruments can be obtained.
• Universality of target gene: Significant results can be obtained regardless of kinds of target genes.

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: Yes
Fragment Size: ~1 kb

Application

- Real-Time Quantification of DNA/cDNA targets.
- Quantification of gene expression
- Microbial/Viral Pathogen Detection.


Figure 1. Real-Time PCR Data of AccuPower® Greenstar™ qPCR PreMix

AccuPower® Greenstar™ PreMix provides at least 7 orders of magnitude in dynamic range (10 fg ~ 10 ng /reaction).
A : Amplification curve. Lambda DNA primers were added into Greenstar™ qPCR PreMix. A series of Lambda DNA positive control diluents were tested.
B : Standard curve. Data were obtained using Exicycler™ 96 Real-Time Quantitative Thermal Block (Bioneer Co.).
C : Melting curve.

Figure 2. Comparison of amplification quality between AccuPower® Greenstar™ qPCR PreMix and other supplier's master mixture.

Amplification & Standard curve of AccuPower® Greenstar™ qPCR PreMix and other supplier’s master mixture.
Lambda DNA primers were added into AccuPower® Greenstar™ qPCR PreMix and other supplier’s master mixture. A series of Lambda DNA positive control diluents were tested. Reaction mixtures were prepared and qPCR was performed according to each supplier’s protocol.
All data were obtained using ABI 7500 Fast Real-Time PCR machine (Applied Biosystems co.).

Notice to Purchaser

FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.

The lyophilized composition of this product is US Patent Nos. 5861251 and 6153412. Certain applications of this product are covered by pending or issued patents in certain countries. Because purchase of this product does not include a license to perform any patented application, users of this product may be required to obtain a patent license depending upon the particular application and country in which the product is used. Products may be covered by pending or issued patents. Please contact us for more information.

Note: 'Top DNA Polymerase' of this product is specifically optimized for increasing base incorporation rate by inactivating 5'->3' exonuclease activity. Therefore, this is not recommended to use for Real Time PCR using Taqman Probe, for which AccuPower® Greenstar™ qPCR PreMix.

Trademark: 'AccuPower®' is a registered trademark of Bioneer Corporation. 'DualStar™' is a trademark of Bioneer Corporation.

* Some of the products may not be available in some countries, For details, please contact your local distributor.

MSDS
• AccuPower® Greenstar™ qPCR PreMix

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

Ready-to-use cocktail containing all components,except primer, for the amplification and detection of DNA in real-time quantitative PCR(qPCR)


The AccuPower® 2X Greenstar qPCR Master Mix is a ready-to-use cocktail containing all components,except primer, for the amplification and detection of DNA in real-time quantitative PCR(qPCR). It combines the automatic "Hotstart" technology of Top DNA polymerase and SYBR Green I fluorescent dye to deliver excellent sensitivity in the quantification of target sequences, with a linear dose response over a wide range of target concentration. Volumes are provided for 100 or 200 amplification reactions of 50ul each.

Features and Benefits

• High Specificity :  AccuPower® 2X Greenstar qPCR Master Mix provides more accurate Real-time PCR result by application of Hot-start method.
• Stability:  The chemical stabilizer maintains enzyme activity for 2 years at -20°C
• Simplicity: Ready to use, AccuPower® 2X Greenstar qPCR Master Mix contains everything of Real-time PCR excluding primer and template.
• Reproducibility:  Bioneer's strict quality controlled production system ensures that your results will be reproducible experiment after experiment

- Real-time quantification of DNA and cDNA targets
- Gene expression profiling
- Microbial & Viral pathogen detection

Experimental Data


 

Fig 1. Highly reproducible Ct values
Amplification of an 90-bp target gene was detected using serially diluted LP(Legionella Pneumoniae) genomic DNA (from 10⁶ copies to 10¹ copies ) with AccuPower® 2X Greenstar qPCR Master mix.
As shown in Fig. Highly reproducible Ct values were achieved within each Lot. set of triplicates

Fig 2. Comparison of the Specificity of the SYBR® Green assay
Amplification of an 90-bp target gene was detected using serially diluted LP(Legionella Pneumoniae) genomic DNA (10n dilution;10⁵~10¹copies) with AccuPower® 2X Greenstar qPCR Master mix.
As shown in Fig. Very small amount of primer dimers was appeared in AccuPower® 2X Greenstar qPCR Master mix than other kits.

Using a Bioneer Exicycler™ 96

- The annealing temperature can be set to 55~65°C, depending on the primer Tm value.
- The annealing time should be set for 5~20 seconds.
  Longer annealing time results in increased efficiency, and a shorter time decreases non- 
  specific amplification.

Fig 3. Gene Expression Analysis
AccuTarget™ Validated Real-Time PCR Primer Library for Human is designed by Bioneer’s bioinformatics tool and targeting for human genome. cDNA was synthesized using Human PTGS2 target primer of those and Human total RNA identically quantified from Hela cell and blood cell with AccuPower® CycleScript RT PreMix(K-2044, Bioneer). Gene analysis was carried out both Hela cell and blood cell by operating Real-time PCR reaction(95°C 10 min, 1 cycle and 95°C 10 sec, 58°C 25 sec, 72°C 30 sec, 41 cycles) using the cDNA, AccuPower® 2X Greenstar qPCR Master Mix and Exicycler™ 96 Real-Time Thermal Block(Cat. No. A-2060).

MSDS
• AccuPower® 2X Greenstar master mix solution

Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

Features and Benefits

• User Convenience: For this you can add probe and primers for your target gene simply. In addition, you can satisfy your result by using this kit for several different types of qPCR machine.
• Dynamic Range: There are wide ranges of copies like 8 log from 10 to 108 copies.
• Specificity: You can get the best amplification result with your target gene by using Hotstart Taq DNA polymerase 
• Universality of target gene: You always achieve great real time PCR result even though you use various types of DNA, cDNA, high GC template and more.
• Easy-to-use: In this kit, we provide the product formation like freeze-dried premix or master mix type included thermostable DNA polymerase, dNTPs and provide separately and individually. Therefore, the users add only Template, Primers and DEPC distilled water when you run your real time PCR.
• Reproducibility: Under ISO 9001 Quality Assurance System, our mass produced product AccuPower® Plus Dualstar™ qPCR PreMix, 2X Master mix give you great satisfactory with uniform amplified effectiveness in all your reaction tube.