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The DNA sequence specificity of hedamycin damage determined by ligation‐mediated PCR and linear amplification
Author(s) -
Cairns Murray J.,
Murray Vincent
Publication year - 1998
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549800203782
Subject(s) - primer (cosmetics) , multiplex ligation dependent probe amplification , ligation , dna sequencing , dna , dna damage , primer extension , microbiology and biotechnology , primer dimer , sequence (biology) , computational biology , multiple displacement amplification , base pair , biology , polymerase chain reaction , genetics , chemistry , base sequence , gene , dna extraction , multiplex polymerase chain reaction , organic chemistry , exon
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.