Development of a new lysis solution for releasing genomic DNA from bacterial cells for DNA amplification by polymerase chain reaction
Abolmaaty, A.; Vu, C.; Oliver, J.; Levin, R.E.
Microbios 101(400): 181-189
2000
ISSN/ISBN: 0026-2633 PMID: 10756522 Document Number: 512801
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 O 157: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 O 157: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.