Use of Multiscreen plates for the preparation of bacterial DNA suitable for PCR.
A rapid and inexpensive method for isolating bacterial DNA for use in PCR is described. The method is based on the guanidinium thiocyanate (GuSCN)-lysis method of Boom et al. (J. Clin. Microbiol. 28:495-503, 1990) and enables a multiple of 96 samples to be prepared in only one hour. We use Multiscreen plates and a vacuum manifold from Millipore. Clinical samples are lysed and washed in the wells of a Multiscreen plate, and DNA is eluted in a standard microplate. Purified DNA was recovered with high yields (over 25%). The method allows multichannel or robotic pipetting for both the sample preparation as well as for the PCR step. The method has been applied successfully to detect pathogenic Streptococcus suis type 2 in nasal and tonsil swab specimens of pigs.
Institute for Animal Science and Health (ID-DLO), Lelystad, The Netherlands.
F H
FH
Reek
M A
MA
Smits
E M
EM
Kamp
H E
HE
Smith
D000818:Animals; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D005260:Female; D006146:Guanidines / Q000737:chemistry; D009297:Nasal Mucosa / Q000382:microbiology; D014066:Palatine Tonsil / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D017011:Streptococcus suis / Q000235:genetics; D013552:Swine; D013861:Thiocyanates / Q000737:chemistry
D004269:DNA, Bacterial; D006146:Guanidines; D013861:Thiocyanates; C054436:guanidine thiocyanate