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

Authors
Iakobashvili, R Lapidot, A
Citation
R. Iakobashvili et A. Lapidot, Low temperature cycled PCR protocol for Klenow fragment of DNA polymerase I in the presence of proline, NUCL ACID R, 27(6), 1999, pp. 1566-1568
Citations number
10
Categorie Soggetti
Biochemistry & Biophysics
Journal title
NUCLEIC ACIDS RESEARCH
ISSN journal
03051048 → ACNP
Volume
27
Issue
6
Year of publication
1999
Pages
1566 - 1568
Database
ISI
SICI code
0305-1048(19990315)27:6<1566:LTCPPF>2.0.ZU;2-#
Abstract
A method for performing cycled PCR at low temperatures, using the thermolab ile Klenow fragment of DNA polymerase I, is reported. Application of prolin e as a buffer additive in the range of 3,0-5.5 M remarkably increases the t hermal stability of the polymerase and decreases the denaturation temperatu re of DNA template, This method might be applicable to a broad spectrum of thermolabile DNA polymerases in cycled PCR and other methods of DNA amplifi cation.