Improved silica-guanidiniumthiocyanate DNA isolation procedure based on selective binding of bovine alpha-casein to silica particles.
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.
Boom
R
R
Laboratory of Medical Microbiology, Department of Virology, Section of Clinical Virology, Academic Medical Center, University of Amsterdam, 1100 DD Amsterdam, The Netherlands.
Sol
C
C
Beld
M
M
Weel
J
J
Goudsmit
J
J
Wertheim-van Dillen
P
P
D000818:Animals; D002364:Caseins / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle; D017931:DNA Primers; D004279:DNA, Viral / Q000134:cerebrospinal fluid / Q000302:isolation & purification* / Q000652:urine; D006146:Guanidines; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D011485:Protein Binding; D015183:Restriction Mapping; D012680:Sensitivity and Specificity; D012822:Silicon Dioxide; D013861:Thiocyanates; D014777:Virus Diseases / Q000134:cerebrospinal fluid / Q000175:diagnosis / Q000652:urine