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DNA Amplification

High-Fidelity PCR

The fidelity of a polymerase refers to its ability to insert the correct base during PCR. Conversely, the rate of misincorporation is known as a polymerase’s error rate. High-fidelity PCR, utilizes a DNA polymerase with a low error rate and results in a high degree of accuracy in the replication of the DNA of interest. NEB scientists were the first to identify and commercialize a high-fidelity DNA polymerase suitable for PCR, namely Vent® DNA Polymerase. Since its discovery, several other high fidelity polymerases have been discovered and engineered – all with differing processivity, speed and levels of accuracy.

When do you need a high-fidelity dna polymerase?

Comparison of High-Fidelity Polymerases

1 Potapov, V., & Ong, J. L. (2017). Examining Sources of Error in PCR by Single_Molecule Sequencing. PloS one, 12(1), e0169774. doi:10.1371/journal.pone.0169774 PMID: 28060945

2 PCR-based mutation screening in lacZ (NEB), lacI (Agilent) or rpsL (Life)

3 Takagi et al (1997) Appl. Env. Microbiol. 63, 4504-4510.

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Protocols for High-Fidelity PCR
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