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
Subjects:
Cell Biology;
Molecular Biology;
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
View Article:
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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.
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| Keywords: Hedamycin; dideoxy sequencing; chemical sequencing; DNA ligase; DNA polymerase |
| view references (22) |

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