Improved Silica-Guanidiniumthiocyanate DNA Isolation Procedure Based on Selective Binding of Bovine Alpha-Casein to Silica Particles

Journal of Clinical Microbiology - Tập 37 Số 3 - Trang 615-619 - 1999
René Boom1, C. J. A. Sol1, Marcel Beld2, Jan Weel1, Jaap Goudsmit2, Pauline Wertheim‐van Dillen1
1Laboratory of Medical Microbiology, Department of Virology, Section of Clinical Virology,1 and
2Department of Retrovirology,2 Academic Medical Center, University of Amsterdam, 1100 DD Amsterdam, The Netherlands

Tóm tắt

ABSTRACT DNA purified from clinical cerebrospinal fluid and urine specimens by a silica-guanidiniumthiocyanate procedure frequently contained an inhibitor(s) of DNA-processing enzymes which may have been introduced by the purification procedure itself. Inhibition could be relieved by the use of a novel lysis buffer containing alpha-casein. When the novel lysis buffer was used, alpha-casein was bound by the silica particles in the first step of the procedure and eluted together with DNA in the last step, after which it exerted its beneficial effects for DNA-processing enzymes. In the present study we have compared the novel lysis buffer with the previously described lysis buffer with respect to double-stranded DNA yield (which was nearly 100%) and the performance of DNA-processing enzymes.

Từ khóa


Tài liệu tham khảo

10.1016/0005-2787(72)90426-1

10.1016/0168-1702(85)90242-4

10.1016/S0167-4838(96)00159-8

10.1002/elps.1150080203

Boom R. Unpublished data.

10.1128/jcm.28.3.495-503.1990

Boom R. et al. Submitted for publication.

10.1016/0140-6736(91)90492-8

10.1016/S0162-0134(96)00147-X

10.1128/jcm.30.12.3185-3189.1992

10.1093/nar/19.15.4295

10.1016/0166-0934(96)02063-0

10.1136/bmj.306.6877.546

10.1016/0166-0934(95)01966-9

10.1007/BF01921720

10.1093/nar/9.13.2989

10.1128/aem.62.3.1102-1106.1996

10.1038/227680a0

10.1017/S0022029900028417

10.1128/jcm.35.3.708-713.1997

10.1126/science.239.4839.487

10.1093/infdis/171.2.273

Swaisgood H. E. Chemistry of the caseins Advanced dairy chemistry Fox P. F. 1 1992 63 110 Elsevier Applied Science London, United Kingdom

Vernazza P. L. Dyer J. R. Fiscus S. A. Eron J. J. Cohen M. S. HIV-1 viral load in blood, semen and saliva.AIDS11199710581059

10.1128/jcm.31.5.1241-1245.1993

10.1016/0166-0934(93)90006-D