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 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.
Abolmaaty
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Department of Food Science, Massachusetts Agricultural Experiment Station, University of Massachusetts, Amherst 01003, USA.
Vu
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Oliver
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Levin
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D002021:Buffers; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification*; D019453:Escherichia coli O157 / Q000737:chemistry* / Q000235:genetics; D005470:Fluorometry; D007091:Image Processing, Computer-Assisted; D007202:Indicators and Reagents / Q000494:pharmacology; D008089:Listeria monocytogenes / Q000737:chemistry / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D016958:Pseudomonas putida / Q000737:chemistry / Q000235:genetics; D012477:Salmonella enteritidis / Q000737:chemistry / Q000235:genetics; D019810:Sodium Azide / Q000494:pharmacology
D002021:Buffers; D004269:DNA, Bacterial; D007202:Indicators and Reagents; D019810:Sodium Azide