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Friday, 01 March 2013 11:15

AccuPower Gold Multiplex PCR PreMix

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AccuPower Gold Multiplex PCR PreMix can amplify up to 20 target genes in a single tube. AccuPower Gold Multiplex PCR PreMix contains Gentaur’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. Gentaur’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.


Specifications

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


Application

Target Application
Human and Animal
  STR analysis for determining genetic profiles in forensic cases
  Molecular diagnostic analysis
  Genotyping assay
  Qualitative and semi-qualitative gene expression assay
  Mutant screening
  Transgenic organism analysis
Plant
  STR analysis
  Detection of pathogens/bacterial infection
  Transgenic organism analysis
  Qualitative and semi-qualitative gene expression assay
 
9CycleScript RT gentaur RT-PCR PreMix RocketScript gentaur Cycle RT antibodies PreMix DualStar qPCR gentaur
Figure 1. 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. The last lane labeled Multi represents all amplicons amplified in a single reaction. 
a) 10-plex PCR, b) 20-plex PCR
 
10elisa pcr gentaur taq PyroHotStart HotStart
Figure 2. 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 AccuPowerGold Multiplex PCR PreMix reactions.
 
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).
 
11antibodies PreMix DualStar qPCR gentaur antibodies PreMix DNA Ligation
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).
 
12premix accupower RocketScript RT PreMix accu power
Figure 4. Comparison of amplification quality, using a Caliper Labchip® 90 system, between AccuPower Gold Multiplex PCR PreMix and competitor Multiplex PCR kits.
a) Virtual Gel Image. Gel image illustrates data reproducibility of the Caliper LabChip® 90 system.
b) Overlay of expression levels using Gentaur’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.

 

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