publication . Article . 2017

Detection of gene mutations in genome “hot” spots: “hairpin” amplicons in DNA melting analysis

V. N. Kondratova; I. V. Botezatu; V. P. Shelepov; Anatoly V. Lichtenstein;
Open Access Russian
  • Published: 01 Apr 2017 Journal: Uspehi Molekulârnoj Onkologii, volume 4, issue 1, pages 46-52 (issn: 2413-3787, Copyright policy)
  • Publisher: ABV-press
Abstract
Polymerase chain reaction (PCR) followed by DNA melting analysis with TaqMan probes effectively reveals mutations in the human genome “hot” spots. The necessity to carry out PCR in the asymmetric variant causes, however, a number of restrictions of this method: 1) an inability of quantitative estimates of gene copy numbers; 2) the need for 2 independent PCR tests for detection of mutations in both complementary strands of an amplicon (this approach improves reliability and sensitivity of the analysis); 3) the complication of PCR design and decrease in efficiency of amplification. Overcoming these restrictions was possible by means of symmetric PCR with primers c...
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free text keywords: mutation scanning, dna melting analysis, taqman probes, kras, “hairpin” amplicons, lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens, lcsh:RC254-282, Biology, Amplicon, Molecular biology, Gene mutation, Genetics, Nucleic acid thermodynamics, Genome
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