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

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 lev els of genomic DNA from Escherichia coli 0157:H7 cells compared with a numb er of other commonly used cell lysis methods. Ethidium bromide stained DNA bands resulting from PCR amplification of target DNA from 100 CFU of E. col i 0157:H7 were readily detected following electrophoresis of agarose gels. In contrast, conventional cell lysis methods failed to detect target DNA fr om 100 CFU after PCR amplification. The new solution was highly effective f or lysing cell suspensions of Salmonella enteritidis, Pseudomonas putida, L ysteria monocytogenes and Psychrobacter immobilis.