Low temperature cycled PCR protocol for Klenow fragment of DNA polymerase I in the presence of proline.
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.
Department of Organic Chemistry, The Weizmann Institute of Science, Rehovot 76100, Israel.
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Iakobashvili
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Lapidot
D004256:DNA Polymerase I / Q000187:drug effects*; D004795:Enzyme Stability / Q000187:drug effects; D009691:Nucleic Acid Denaturation / Q000187:drug effects; D016133:Polymerase Chain Reaction / Q000379:methods*; D011392:Proline / Q000494:pharmacology*
D011392:Proline; D004256:DNA Polymerase I