Low Temperature Cycled PCR Protocol for Klenow Fragment of DNA Polymerase I in the Presence of Proline

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1999 Feb 26

PMID 10037825

Citations 5

Authors

R Iakobashvili

A Lapidot

Affiliations

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Abstract

A method for performing cycled PCR at low temperatures, using the thermolabile Klenow fragment of DNA polymerase I, is reported. Application of proline as a buffer additive in the range of 3.0-5.5 M remarkably increases the thermal stability of the polymerase and decreases the denaturation temperature of DNAtemplate. This method might be applicable to a broad spectrum of thermolabile DNA polymerases in cycled PCR and other methods of DNA amplification.

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