ExiProgen™

    

Bioneer’s ExiProgen™ is a breakthrough in synthetic biology allowing for the synthesis and purification of one to 16 proteins per run. DNA, in the form of plasmid or linear PCR product, is added to the system and Bioneer’s coupled transcription/translation E. coli extract then synthesizes the protein. The crude protein extract is further purified, using His-Tag engineered into the proteins. The net result is up to 100 ug of >90% pure Protein in about 6 hours. ExiProgen™ can also purify Nucleic Acids from several sources, making it one of the most versatile systems available.

Features and Benefits

• Fast Protein production : From DNA to Pure protein in about 6 hours
• DNA purification : A wide variety of gDNA kits available
• RNA purification : A wide variety of RNA kits available
• Affinity protein purification : For couples expression and purification in one system!

Built in Protocols optimized for protein synthesis/purification and the extraction of a wide variety of Nucleic Acid Samples ExiProgen™ has more contains over 900 protocols, each optimized for protein synthesis/purification and target nucleic acid type and source sample. This optimization enables the user to obtain reproducible results for every run, every day. The instrument software can also be upgraded through the network connection port so you can stay up-to-date with the best performing protocols
Cooling Block ExiProgen™ Has a built-in cooling block where the elution tube rack sits. Sample integrity is ensured by keeping the samples below 10°C. This allows for overnight runs and provides you with confidence in your results
Magnetic block & Heating block To increase extraction efficiency, the ExiProgen™ has an integrated combined magnetic/heating block. The combination of bead magnetization and sample heating reduces experiment time and increases elution efficiency, resulting in increased sample yield. The heating block’s precision temperature control also ensures reproducible results for protein synthesis reactions
Contamination Shield ExiProgen™ comes with a contamination shield designed to protect the assay from cross-contamination during instrument operation. Any time the pipette tips are moving, the contamination shield will slide under the tips, therefore eliminating the possibility of intra-assay cross-contamination which is a must when working with multiple samples
Easy to use with touch screen The 3.5" touchscreen maximizes efficiency by offering an intuitive interface with simple push-button operation for processes such as selecting protocols and controlling the UV sterilization lamp.
UV lamp ExiProgen™ has a powerful UV sterilization lamp that enables the user to sterilize the instrument chamber before and/or after every nucleic acid extraction or protein purification run. This prevents possible inter-assay cross-contamination that may occur on a busy work day

Experimental Procedure

Principle of protein synthesis and purification

ExiProgen™ Protein Synthesis Kit useds a E. coli extract to effect coupled transcription/translation of input DNA, which can be plasmid, or PCR generated DNA. The protein itself is generated with a His-Tag, which is then purified using the Ni-NTA magnetic bead provided. The result is high yields of protein that is >90% pure.

Nucleic acid extraction principle

ExiProgen™ DNA/ RNA Kits work on the principle of cell lysis, followed by bind, wash elute from silica magnetic beads. High yields of ultrapure DNA or RNA are obtained with OD₂₆₀ readings of > 1.8 for DNA and 2.0 for RNA.

Applications

Enzyme engineering, protein evolution, Synthetic biology, Bio-energy R&D, Protein drug R&D, expression screening, functional analysis, assay development and antigen production.


Specifications

Functions

Experimental Data

1. Protein expression and purification
- Amount of input DNA: 6 - 10 ug (purity >1.8)
- Protocol number : 902
- Total prep time : < 6 h
- Yield: ~100 ug

Procedure




SDS-PAGE analysis

	Figure 1. GFP protein (His-tagged protein)’s expression/purification with ExiProgen™. M; AccuLadder™ Protein Size Maker (Low) E; expression sample P; purification sample
	Figure 2. Various His-tagged proteins expressed/purified with ExiProgen™. M; AccuLadder™ Protein Size Maker (Low) E; expression sample P; purification sample

2. Total RNA Extraction from Tissue or cultured mammalian cell
- Sample: 15 ㎎ (Rat liver tissue) or 1 X 10⁶ cells (HeLa)
- Protocol number: 202
- Average yield: 25 ug (Rat liver tissue), 10 ug (HeLa)
- Average RQS: > 9.5

* RQS(RNA quality score): he Caliper RQS is a calculated score that rates the quality of RNA samples. The RQS has been validated to correlate well with Agilent’s RIN (RNA Integrity Number) and follows the same 0-10 scale rating. Results comparing RIN to RQS for the same samples run on both LabChip GX and Agilent’s Bioanalyzer 2100 typically show <10% deviation. ( http://www.caliperls.jp/assets/pdf/lcgx/lcgx-AP-402.pdf)

Procedure



Rat total RNA Extracted from tissue

Total RNA from Rat was observed to contain intact rRNA with no evidence of degradation. 
 

Real-Time PCR carried out on Rat Total RNA
Real time RT-PCR was carried out.AccuPower® RocketScrpt™ RT PreMix(K-2101, Bioneer), and AccuPower® DualStar™ qPCR PreMix(K-6110, Bioneer) , and the Exicycler™ 96 Real-Time PCR system (A-2060).     


Analysis of total RNA from HeLa cells.

Total RNA was extracted from HeLa cell (1 X 10⁶ cells) and analyzed using the LabChip GX (Caliper Life Science).
Each subunit of rRNA is clearly seen with no detectable degradation . In addition, the average yield of Total RNA was 10 ug, and the average RQS was 9.5 or higher.


Real-Time PCR carried out on total RNA from Hela cells.

Real time RT-PCR was carried out on total RNA extracted from HeLa cell (1 X 10⁶ cells) using AccuPower® RocketScrpt™ RT PreMix(K-2101, Bioneer), AccuPower® DualStar™ qPCR PreMix(K-6110, Bioneer) and the Exicycler™ 96 Real-Time PCR system(A-2060)

 

3. Genomic DNA from Cultured mammalian cell (HeLa))
- Sample volume: 1 X 10⁶ cells
- Protocol number: 109
- Total prep time: < 1hr
- Average yield: 4-8 ug

Procedure



Agarose gel analysisAgarose gel analysis

Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 1X10⁶ cells of E.coli cells and Lanes 2, 4, 6, 8, 10, 12, 14, 16 were extracted with ddH₂O as a negative control in DNA extraction. Note all samples have very similar yields.
Purity was also tested and was consistently between 1.9 and 2.0 (not shown).

 

4. Genomic DNA from Tissue (Bovine skeletal muscle)
- Sample volume: 1 X 10⁶ cells
- Protocol number: 109
- Total prep time: < 1hr
- Average yield: 8-12 ug

Procedure


Agarose gel analysis

Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 20 mg of bovine tissues and Lanes 2, 4, 6, 8, 10, 12, 14, 16 were extracted with ddH₂O as a negative control in DNA extraction. Note all samples have very similar yields.
Purity was also tested and was consistently between 1.9 and 2.0 (not shown).

 

5. Genomic DNA from Plant Tissue (Spinach leaf)
- Sample volume: 100 ㎎
- Protocol number: 104
- Total prep time: <1 hr
- Average yield: 3-5 ug

Procedure

Agarose gel analysis

Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 100 mg of spinach leaves and Lanes 2, 4, 6, 8, 10, 12, 14, 16 were extracted with ddH₂O as a negative control in DNA extraction. Note all samples have very similar yields.
Purity was also tested and was consistently between 1.9 and 2.0 (not shown).Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 100 mg of spinach leaves and Lanes 2, 4, 6, 8, 10, 12, 14, 16 were extracted with ddH₂O as a negative control in DNA extraction. Note all samples have very similar yields.
Purity was also tested and was consistently between 1.9 and 2.0 (not shown).

 

 

 

Application Note

Protein Synthesis

• ExiProgen™_Application_Fluorescent proteins

• ExiProgen™_Application_High molecular weight protein

• ExiProgen™_Application_in vivo protein expression pET vectors

Manual

• ExiProgen™

Brochure

• ExiProgen™

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

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A-5041	ExiProgen™	Inquire