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 microcolumns 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 а few copies of DNA.
Download PDF:
Citation:
Fedchenko V.I., Guriev S.O., Kaloshin A.A. (2004) Effective method of DNA purification from biological and non-biological contaminations and optimization of its amplification. Biomeditsinskaya Khimiya, 50(2), 211-216.
Fedchenko V.I. et al. Effective method of DNA purification from biological and non-biological contaminations and optimization of its amplification // Biomeditsinskaya Khimiya. - 2004. - V. 50. -N 2. - P. 211-216.
Fedchenko V.I. et al., "Effective method of DNA purification from biological and non-biological contaminations and optimization of its amplification." Biomeditsinskaya Khimiya 50.2 (2004): 211-216.
Fedchenko, V. I., Guriev, S. O., Kaloshin, , A. A. (2004). Effective method of DNA purification from biological and non-biological contaminations and optimization of its amplification. Biomeditsinskaya Khimiya, 50(2), 211-216.
References
Midler S.A., Dykes D.D. and Polesky H.F. (1982) Nucl. Acids Res.. 16, 1215- 1223. Scholar google search
Lin D., Blom B. and Flolmlund G. (1988) Gene Anal. Techn., 5, 97- 101. Scholar google search
Gustincich S., Manfioletti G., Del Sal G., Carninci P. (1991) BioTechniques., 11, 298- 302. Scholar google search
Roos J.A., Nelson G.B., Holden K.L., (1992) Nucl. Acids Res., 19, 6053- 6054. Scholar google search
Zeillinger R., Schneeberger C., Speiser P., Kury F. (1993) Bio- Techniques., 14, 202- 203. Scholar google search
Ефремов И.А., Носиков В.В., Скоблилов Е.Ю., Законова А.Ф., Иванов П.Л. (2001) Судебно- медицинская экспертиза, 44, 11- 17. Scholar google search
Comay C.T., Koons В. W., Presley К.W., Smerick J.B., Sobciralski C.A., Stanley D.M., Bacchtel F.B. (1994) J. Forensic Sci., 39, 1254- 1269. Scholar google search
Hochmeister M.N., Budowle В., Borer U.V., Eggmann U., Сотеу С Т., Dirnholer R. (1991) J. Forensic Sci., 36, 1649- 1661. Scholar google search
Lahiri D.K., Bye S., Nurnberger J.I., Hades M.E., Crisp M. (1992) J. of Biochem. Biophys. Meth., 25, 193- 205. Scholar google search
Honda K., Nakatome M., Islam M.M., Bai H., Ogura Y., Kuroki H., Yamazaki M., Terada M., Misawa S., Wakasugi С. (1995) Forensic Sci., 40, 637- 640. Scholar google search
Schmitl C., Schmutzler A., Prim M., Staak M. (1994) Forensic Sci Int.. 66, 129- 41. Scholar google search
Maniatis Т., Fritach E.F andSambrook (1993) J. Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY. Scholar google search
Boerwinkle E., Xiong W., Fourest E. and Chan L. (1989) Proc. Natl. Acad. Sci. USA, 86, 212- 216. Scholar google search
G. Т., Richards В., and Klinger K. W. (1989) Nucl. Acids Res., 17, 2140. Scholar google search
Hagelberg E., Sykes В., and Hedges R. (1989) Nature. 342, 485. Scholar google search
Singer- Sam J., Tanguay R.L., Riggs A. (1989) Amplifications, 3, 11. Scholar google search
Walsh P.S., Metzger D.A., Higuchi R. (1991) Bio- Techniques, 10, 506- 513. Scholar google search
