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Journal of Clinical Microbiology, March 1999, p. 615-619, Vol. 37, No. 3
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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

René Boom,1,* Cees Sol,1 Marcel Beld,2 Jan Weel,1 Jaap Goudsmit,2 and Pauline Wertheim-van Dillen1

Laboratory of Medical Microbiology, Department of Virology, Section of Clinical Virology,1 and Department of Retrovirology,2 Academic Medical Center, University of Amsterdam, 1100 DD Amsterdam, The Netherlands

Received 15 April 1998/Returned for modification 23 June 1998/Accepted 3 December 1998

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.

* Corresponding author. Mailing address: Laboratory of Medical Microbiology, Department of Virology, Section of Clinical Virology, Meibergdreef 9, 1100 DD Amsterdam, The Netherlands. Phone: (31-20)-5665472. Fax: (31-20)-6974005.

Journal of Clinical Microbiology, March 1999, p. 615-619, Vol. 37, No. 3
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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