High fidelity PCR demands a low rate of nucleotide misincorporation (mismatch). Sequence integrity is particularly important in applications involving gene expression, low-copy-number templates, long templates, cDNA library construction, SNP analysis, and site-directed mutagenesis.
How is it achieved?
The intrinsic properties of polymerases, such as extension rates and 3'-5' exonuclease (proofreading) activity directly influence fidelity. In practice, a mixture of high yield but low fidelity polymerases (i.e. Taq), with high fidelity polymerases (i.e. pfu, Vent) can enhance overall amplification performance. External factors like template sequence, buffer composition, and reaction conditions are also important for high fidelity PCR.
What makes our PCR polymerases the best?
Our high performance enzyme mixes and kits are optimized for a variety of high fidelity PCR applications.
Advantage HF 2 PCR Kit combines Advantage 2 with a proprietary mix of dNTPs and a specially formulated buffer that work together to achieve 30-fold higher fidelity than wild-type Taq DNA Polymerase.
Advantage HD Polymerase is a novel PCR enzyme with a robust 3'–5' exonuclease activity that provides superb accuracy, with a low error rate of 1 error per 20 kb (error rate=# misincorporated bases/# bases synthesized).