Application of Repair Enzymes to Improve the Quality of Degraded DNA Templates for PCR Amplification

作者:Dovgerd A P*; Zharkov D O
来源:APPLIED BIOCHEMISTRY AND MICROBIOLOGY, 2014, 50(3): 235-242.
DOI:10.1134/S0003683814030053

摘要

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 modeldamaged substrates with this set of enzymes resulted in an increased PCR product yield as compared with that of the unrepaired samples.

  • 出版日期2014-5

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