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
UDG and dUTP in the MasterMix prevent the re-amplification 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
• Specificity
Pyrophosphate (PPi) has high affinity for Mg2+. By adding PPi to the reaction mixture, the 2+ 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.
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