PCR is used for the amplification of a next-generation sequencing (NGS) library. In addition to increasing the amount of DNA, the PCR amplification step is also used for DNA target enrichment for fragments that have an adaptor-ligated to each end. These will be the fragments that attach to the flow cell and will be sequenced, so it is important that this PCR amplification step does not introduce mutations to the NGS library, as a lower error rate means that less sequence coverage is required, saving expense and time.

High-Fidelity Pfu is a thermostable high fidelity DNA polymerase enzyme, isolated from Pyrococcus furiosus. The Pfu polymerase enzyme replicates DNA in the 5´→3´ direction in the presence of magnesium, but also possesses 3´→5´ exonuclease (proofreading) activity. Base misincorporations that may occur during polymerization are rapidly excised by this proofreading activity. High-Fidelity has an error rate of 3.0 x 10-6 generating blunt-ended amplicons up to 5 kb in length. Glycerol-Free High-Fidelity Pfu (HC) is supplied in a glycerol-free storage buffer and is accompanied by a 5x Reaction Buffer that contains magnesium, dNTPs and excipients required for lyophilisation.

• High concentration Pfu polymerase allowing for bulk production of PCR master mixes
• An error rate of 3.0 x 10-6
• Ideal for NGS library amplification
• Glycerol-free for preparation of dried-down PCR tests