Development of a new lysis solution for releasing genomic DNA from bacterial cells for DNA amplification by polymerase chain reaction.

Microbios Pub Date : 2000-01-01
A Abolmaaty, C Vu, J Oliver, R E Levin
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Abstract

A new lysis solution designated TZ, consisting of 2.0% Triton X-100 plus 2.5 mg sodium azide/ml in 0.1 M Tris-HCl buffer at pH 8.0, yielded higher levels of genomic DNA from Escherichia coli O157:H7 cells compared with a number of other commonly used cell lysis methods. Ethidium bromide stained DNA bands resulting from PCR amplification of target DNA from 100 CFU of E. coli O157:H7 were readily detected following electrophoresis of agarose gels. In contrast, conventional cell lysis methods failed to detect target DNA from 100 CFU after PCR amplification. The new solution was highly effective for lysing cell suspensions of Salmonella enteritidis, Pseudomonas putida, Lysteria monocytogenes and Psychrobacter immobilis.

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从细菌细胞中释放基因组DNA用聚合酶链反应扩增的新型裂解液的研制。
一种新的裂解溶液TZ,由2.0% Triton X-100和2.5 mg叠氮化钠/ml组成,在0.1 M Tris-HCl缓冲液中,pH为8.0,与许多其他常用的细胞裂解方法相比,从大肠杆菌O157:H7细胞中产生更高水平的基因组DNA。对大肠杆菌O157:H7 100 CFU的靶DNA进行PCR扩增,得到溴化乙啶染色的DNA条带,琼脂糖凝胶电泳后很容易检测到。相比之下,传统的细胞裂解方法无法检测到PCR扩增后100 CFU的目标DNA。该溶液对肠炎沙门氏菌、恶臭假单胞菌、单核增生李斯特菌和固定化冻杆菌的细胞悬浮液有较好的裂解效果。
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