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[Application of repair enzymes to improve the quality of degraded DNA templates for PCR amplification].

Authors :
Dovgerd AP
Zharkov DO
Source :
Prikladnaia biokhimiia i mikrobiologiia [Prikl Biokhim Mikrobiol] 2014 May-Jun; Vol. 50 (3), pp. 264-72.
Publication Year :
2014

Abstract

PCR amplification of severely degraded DNA, including ancient DNA, forensic samples, and preparations from deeply processed foodstuffs, is a serious problem. Living organisms have a set of enzymes to repair lesions in their DNA. In this work, we have developed and characterized model systems of degraded high-molecular-weight DNA with a predominance of different types of damage. It was shown that depurination and oxidation of the model plasmid DNA template led to a decrease in the PCR efficiency. A set of enzymes performing a full cycle of excision repair of some lesions was determined. The treatment of model-damaged substrates with this set of enzymes resulted in an increased PCR product yield as compared with that of the unrepaired samples.

Details

Language :
Russian
ISSN :
0555-1099
Volume :
50
Issue :
3
Database :
MEDLINE
Journal :
Prikladnaia biokhimiia i mikrobiologiia
Publication Type :
Academic Journal
Accession number :
25757334
Full Text :
https://doi.org/10.7868/s0555109914030210