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[Application of repair enzymes to improve the quality of degraded DNA templates for PCR amplification].
- 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.
- Subjects :
- Animals
Bacteriophage T4 chemistry
Cattle
DNA Damage
DNA Glycosylases chemistry
DNA Ligase ATP
DNA Ligases chemistry
DNA-(Apurinic or Apyrimidinic Site) Lyase chemistry
Escherichia coli chemistry
Humans
Pancreas chemistry
Pancreas enzymology
Polynucleotide 5'-Hydroxyl-Kinase chemistry
Ribonucleases chemistry
Taq Polymerase chemistry
Bacterial Proteins chemistry
DNA Repair
DNA, Bacterial chemistry
Plasmids chemistry
Polymerase Chain Reaction methods
Viral Proteins chemistry
Subjects
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