AccuPower® 2X Greenstar™ qPCR Master Mix


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
]]> Applications

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

 

Experimental Data


 

Step Condition Cycle
Pre-Denaturation 95°C, 15min 1
Denaturation 95°C, 15sec 45
Annedling/Extension 60°C, 30sec
Detection(Scan)  
Melting 65°C~90°C, every 1°C, 1sec 1

 

PCR reaction mixture Perreaction
2X Greenstar Master Mix 25ul
PCR F-Primer (10 pmole) 1.5ul
PCR R-Primer (10 pmole) 1.5ul
Template 5ul
DEPC-distilledwater. A djust to 50ul

 

Copy C(t) Value
Batch 1 Batch 2 Batch 3 Error Rate
NTC UD UD UD 0.5 C(t)
10 36.33 36.74 35.54
100 32.22 31.77 31.73
1000 28.51 28.7 28.04
10000 25.09 25.12 24.73
100000 21.27 21.43 21.47
1000000 18.12 18.13 18.04
10000000 14.99 14.43 14.45

 

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.


 

Target Gene hGAPDH hPTGS2
Sample No. 1 2 1 2
C(t) Value 23.73 23.46 33.98 30.52

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).

 

]]> Manual
• AccuPower® 2X Greenstar master mix solution


MSDS
• AccuPower® 2X Greenstar master mix solution


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

 

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