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. 1999 Mar 15;27(6):1566–1568. doi: 10.1093/nar/27.6.1566

Low temperature cycled PCR protocol for Klenow fragment of DNA polymerase I in the presence of proline.

R Iakobashvili 1, A Lapidot 1
PMCID: PMC148357  PMID: 10037825

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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