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Friday, 01 March 2013 13:33

AccuPower DualStar qPCR PreMix

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

DualStar qPCR PreMix eliminates nonspecific reaction, while providing high sensitivity, due to our novel HotStart methodology. Gentaur uses a unique enzyme-mediated HotStart PCR that provides robust and reproducible results. Gentaur’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.

DualStar Plus qPCR PreMix goes a step further by providing resistance to common inhibitors of PCR (Blood-EDTA, Hemoglobin, Humic Acid from various sources, etc.) providing you with robust and reliable results, even with poor quality samples. Tested for resistance against many common inhibitors of PCR, DualStar plus is the enzyme of choice for any PCR that you want to work right – the first time and every time!

 

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
Reproducibility: Each batch is produced under strict ISO quality controls.
DualStar Plus qPCR PreMix: Available for less samples that will not work in other systems due to low purity

Specifications

5' to 3' exonuclease activity: Yes
3' to 5' exonuclease activity: No
Terminal transferase activity: Yes
Fragment Size: 1 kb


Application

- Gene expression profiling
- Target DNA quantification
- Microbial detection
- SNP analysis
- Viral/bacterial pathogen load determination
- Evaluation of primer pair performance for probe-based real-time PCR

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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 ~ 101copies/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 PCR System (Gentaur).

16elisa pcr gentaur taq PyroHotStart HotStart

Figure 2. Data using Various kinds of Real-time PCR Instruments
AccuPower® DualStar™ qPCR PreMix provides at least 7 orders of magnitude in dynamic range (107 ~ 101copies/reaction).
A: Amplification curve. and standard curve using ABI 7500 Format Fast Real-time PCR machine (Applied Biosystems). A series of WNV positive control diluents were tested.
B: Amplification curve and standard curve using ABI 7500 Format Real-time PCR machine (Applied Biosystems). 
C: Amplification curve and standard curve using Opticon Real-time PCR machine (MJ Research, currently Bio-Rad).

17RT gentaur RT-PCR PreMix RocketScript gentaur Cycle RT

Figure 3. Comparison of detection sensitivity between AccuPower® DualStar™ qPCR PreMix and other supplier's master mixture
West Nile Virus (WNV) primers and TaqMan-based probe were added into AccuPower® DualStar™ qPCR Premix and other supplier's master mixture. A series of WNV 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 Format Fast Real-time PCR machine (Applied Biosystems).

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