PCR amplification of streptococcal DNA using crude cell lysates.
Gram-positive organisms such as streptococci and enterococci are often difficult to lyse. Obtaining DNA for procedures such as PCR amplification usually requires a large scale isolation for each strain under investigation. We describe a simple procedure for small volumes of whole cells, involving pretreatment with detergent and proteinase that allows for efficient release of DNA for PCR amplification. This procedure is fast, reproducible, can be used with a large number of samples, and has been successfully applied to a variety of streptococcal and enterococcal strains.
Hynes
W L
WL
Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City 73190.
Ferretti
J J
JJ
Gilmore
M S
MS
Segarra
R A
RA
D001483:Base Sequence; D002458:Cell Fractionation / Q000379:methods; D004269:DNA, Bacterial / Q000302:isolation & purification*; D003902:Detergents / Q000494:pharmacology; D010450:Endopeptidases / Q000494:pharmacology; D013293:Enterococcus faecalis / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D013297:Streptococcus pyogenes / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development
D004269:DNA, Bacterial; D003902:Detergents; D010450:Endopeptidases