Fidelity of DNA polymerases in DNA amplification.

Fidelity of DNA polymerases in DNA amplification.

Denaturing gradient gel electrophoresis (DGGE) was used to separate and isolate the products of DNA amplification by polymerase chain reaction (PCR). The strategy permitted direct enumeration and identification of point mutations created by T4, modified T7, Klenow fragment of polymerase I, and Thermus aquaticus (Taq) DNA polymerases. Incorrectly synthesized sequences were separated from the wil...

Low-fidelity DNA polymerases

Also known as.... Translesion synthesis polymerases. There are two of them in Escherichia coli, polymerases IV and V, and five in human cells, polymerases ζ, η, ι, κ and Rev1 (so a knowledge of the Greek alphabet is essential when discussing them). Except for polymerase ζ, they fall into a new DNA polymerase family, designated the Y-family. Depending on the lesion, polymerase and context, they ...

Amplification efficiency of thermostable DNA polymerases.

The amplification efficiencies of several polymerase chain reaction (PCR) enzymes were compared using real-time quantitative PCR with SYBR Green I detection. Amplification data collected during the exponential phase of PCR are highly reproducible, and PCR enzyme performance comparisons based upon efficiency measurements are considerably more accurate than those based on endpoint analysis. DNA p...

A reexamination of the nucleotide incorporation fidelity of DNA polymerases.

Intensive study has been devoted to understanding the kinetic and structural bases underlying the exceptionally high fidelity (low error frequencies) of the typical DNA polymerase. Commonly proposed explanations have included (i) the concept of fidelity check points, in which the correctness of a nascent base pair match is tested at multiple points along the reaction pathway, and (ii) an induce...

Regulation and Fidelity: A study of RNA and DNA polymerases