[Effective method of DNA purification from contaminated matters and optimization of amplification].
A new modification of DNA purification has been developed. It includes: 1) standard treatment of biological material with proteinase K followed by phenol-chlorophorm extraction; 2) subsequent sample purification using micro-columns packed with Dowex-50 and Sephadex G-50. Oligonucleotide primers often used for DNA typing in man by means of polymerase chain reaction have also been modified. These are VNTR (variable number of tandem repeats) loci of apoB and D17S5. The increase of stability and specificity of amplification of VNTR loci of apoB and D17S5 was achieved by increase of primer length and amplification cycle. The sensitivity of this mode of amplification is 2-4 ng DNA-template. Employment of the nested amplification for apoB locus increased sensitivity of this method up to a few copies of DNA.
Fedchenko
V I
VI
Orekhovich Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121, Russia. fedval@ibmh.msk.su
Gur'ev
S O
SO
Kaloshin
A A
AA
D001055:Apolipoproteins B / Q000235:genetics; D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification*; D005554:Forensic Medicine; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D020080:Tandem Repeat Sequences
D001055:Apolipoproteins B; D004247:DNA