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The DNA sequence specificity of hedamycin damage determined by ligation-mediated PCR and linear amplification 

Authors: Murray J. Cairns a; Vincent Murray a
Affiliation:   a School of Biochemistry and Molecular Genetics, The University of New South Wales, Sydney, Australia
DOI: 10.1080/15216549800203782
Publication Frequency: 12 issues per year
Published in: journal IUBMB Life, Volume 46, Issue 2 October 1998 , pages 267 - 275
Formats available: PDF (English)
Previously published as: Biochemistry and Molecular Biology International (1039-9712) until 1998

The circumstances under which this title is published have changed:

Reason for change: Changed publisher
Now published by: Wiley-Blackwell Publishing Ltd
Date of change: 2008

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Abstract

The DNA sequence specificity of hedamycin damage was compared using two techniques: Ligation-mediated PCR (LMPCR) and Linear Amplification (LA). The electrophoretic mobility of LA products were made comparable with LMPCR products by using a primer with a 27 bp non-binding 5'-extension. By direct comparison of the damage intensity (as represented by each of the methods), considerable bias was found in the LMPCR system with respect to LA-resulting in the under representation of lesions with T as the base 3' to the damage site. In view of this observation some caution should be exercised in the interpretation of data for DNA damage/repair specificity derived by LMPCR. In addition the extended primer LA method used in these experiments, could be applied to generate a dideoxy sequencing ladder for analysis of LMPCR products. This would negate the need to prepare Maxam-Gilbert chemical sequencing fragments for amplification through LMPCR.
Keywords: Hedamycin; dideoxy sequencing; chemical sequencing; DNA ligase; DNA polymerase
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