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Combine High Yield, Great Accuracy and the,,,Prevention of Carry-over Contamination by Using,,,the Novel Expand High FidelityPLUS PCR System

Roche Applied Science pioneered the blending of thermostable DNA polymerases and the associated buffer systems. The well-known Expand High Fidelity and Expand Long Template PCR Systems were the first commercially available blends to provide amplification of fragments up to 5kb and 20 kb, respectively. The Expand High FidelityPLUS PCR System is a next-generation blend consisting of Taq DNA Polymerase and a novel proofreading protein, isolated and characterized by Roche Applied Science. This protein mediates proofreading activity but has no polymerase activity itself.

The synergy between the mechanism of the proofreading protein and the processivity of Taq DNA Polymerase is the key to Expand High FidelityPLUS Enzyme Blends ability to
  • deliver high yield combined with improved accuracy
  • incorporate dUTP to prevent carry-over contamination when performing amplifications up to 5 kb.
The combination of these characteristics makes the Expand HiFiPLUS Enzyme Blend the product of choice for the simultaneous amplification of a larger number of targets.

Various thermostable DNA polymerases as individual enzymes or blends offer good yield or accuracy. However, the Expand HiFiPLUS Enzyme Blend is the only formulation that delivers high yield and great fidelity for amplification of fragments up to 5 kb.

Expand the Performance of PCRs up to 5 kb

Expand HiFiPLUS Enzyme Blend successfully challenged four products from different suppliers, and proved to be the blend delivering the best sensitivity and specificity for the amplification of fragments up to 5 kb (Figure). Vari ous amounts (in ng) of human genomic DNA were used to amplify a 4.8 kb fragment from the tissue plasminogen activator (tPA) gene. The manufacturers recommended reaction conditions were strictly followed. The result shows that Expand HiFiPLUS produced the best sensitivity, specificity, and yield, even from as little as 1 ng human genomic DNA.



Avoid Cross-Contamination and Obtain Reliable Results

Cross-contamination is a common problem in every lab. Until now, carry-over prevention via the incorporation of dUTP and subsequent treatment with Uracil-DNA Glycosylase was only possible with Taq DNA Polymerase for fragments up to 2.5 kb. The Expand HiFiPLUS Enzyme blend now enables the incorporation of dUTP in fragments up to 5 kb. This makes Expand HiFiPLUS the enzyme of choice for high-fidelity, highyield amplification of longer fragments.



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