Open AccessPCR amplification of streptococcal DNA using crude cell lysates
Author(s) -
Hynes Wayne L.,
Ferretti Joseph J.,
Gilmore Michael S.,
Segarra Robert A.
Publication year - 1992
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1992.tb05303.x
Subject(s) - lysis , dna , polymerase chain reaction , isolation (microbiology) , dna extraction , biology , multiple displacement amplification , microbiology and biotechnology , strain (injury) , proteinase k , genetics , gene , anatomy
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.