Abstract
A method of DNA isolation and purification from highly contaminated sources usual for forensic medicine and criminalistic practice was developed. Oligonucleotide primers were synthesized and the conditions of PCR amplification were optimized for two human minisatellite (VNRT) loci — apolipoprotein B and D1S30 widely used in PCR-based DNA typing. The proposed procedure of DNA isolation includes standard treatment with proteinase K followed by phenol-chloroform extraction and further purification from cations and low molecular weight compounds on a Dowex/Sephadex G-50 minicolumn. The length of the primers and optimal PCR conditions determine stability and specificity of amplification of the VNTR-loci. The sensitivity of the proposed method is 2–4 ng DNA template in the reaction mixture.
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Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 127, No 3, pp. 357–360, March, 1999
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Gur'ev, S.O., Fedchenko, V.I. & Zhdanov, R.I. New method of PCR amplification of two human minisatellite loci and reliable method of DNA isolation. Bull Exp Biol Med 127, 324–327 (1999). https://doi.org/10.1007/BF02433371
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DOI: https://doi.org/10.1007/BF02433371