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Vol 49(2015) N 4 p. 560-568; DOI 10.1134/S0026893315040056 Full Text

R.R. Garafutdinov*, A.A. Galimova, A.R. Sakhabutdinova, V.A. Vakhitov, A.V. Chemeris

DNA amplification using PCR with abutting primers

Institute of Biochemistry and Genetics Ufa Science Center, Russian Academy of Sciences, Ufa, 450054 Russia

*garafutdinovr@gmail.com
Received - 2014-12-18; Accepted - 2015-01-26

DNA analysis of ñîmplex 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.

amplification, PCR, degraded DNA, abutting primers, specificity



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