4.2 Article

DNA amplification using PCR with abutting primers

Journal

MOLECULAR BIOLOGY
Volume 49, Issue 4, Pages 560-568

Publisher

MAIK NAUKA/INTERPERIODICA/SPRINGER
DOI: 10.1134/S0026893315040056

Keywords

amplification; PCR; degraded DNA; abutting primers; specificity

Funding

  1. Department of Biochemical Methods and Nanobiotechnology of the Regional Common Use Center Agidel [NSh-5923.2014.4]

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DNA analysis of A (R) mplex biological objects (wastewater, soil, archaeological and forensic samples, etc.) is currently of great interest. DNA of these objects is characterized by low suitability for research due to the violation of its integrity and chemical structure; thus, the detection of specific nucleic acid fragments can be achieved by PCR with contiguous primers. In this paper, we present the results that clarify the specific characteristics of PCR with abutting primers. The 3'-ends of these primers are annealed at adjacent nucleotides of complementary chains of DNA target. It has been shown that the proximity of primers enables the formation of specific reaction products with a higher sensitivity and less reaction time. Using artificially damaged DNA and DNA from the soil we demonstrated that the abutting primers provide assured detection of specific DNA fragments. The results of this work may be taken into account in PCR with degraded (fragmented) DNA.

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