Reek F H, Smits M A, Kamp E M, Smith H E
Institute for Animal Science and Health (ID-DLO), Lelystad, The Netherlands.
Biotechniques. 1995 Aug;19(2):282-5.
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.
本文描述了一种快速且廉价的用于PCR的细菌DNA分离方法。该方法基于Boom等人（《临床微生物学杂志》28:495 - 503，1990年）的硫氰酸胍（GuSCN）裂解方法，仅需一小时就能制备96个样本的倍数。我们使用密理博公司的Multiscreen板和真空歧管。临床样本在Multiscreen板的孔中进行裂解和洗涤，DNA在标准微孔板中洗脱。纯化的DNA以高产率（超过25%）回收。该方法允许在样本制备以及PCR步骤中使用多通道或机器人移液。该方法已成功应用于检测猪鼻拭子和扁桃体拭子样本中的致病性2型猪链球菌。
