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PCR products generated from unpurified Salmonella DNA are degraded by thermostable nuclease activity
Author(s) -
Gibson J. R.,
McKee R. A.
Publication year - 1993
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.1993.tb00342.x
Subject(s) - nuclease , dna , salmonella , biology , oligonucleotide , microbiology and biotechnology , polymerase chain reaction , palindromic sequence , hot start pcr , chemistry , biochemistry , bacteria , palindrome , genetics , gene , genome , nested polymerase chain reaction
Oligonucleotide primers designed from repetitive extragenic palindromic (REP) sequences were used to PCR‐amplify Salmonella DNA. Unpurified template DNA, present in crude cell extracts, yielded an essentially identical banding pattern to that arising from the use of purified DNA. However, the PCR product derived from the crude preparations did not survive storage at 4°C. This post‐PCR DNA degradation, attributed to endogenous Salmonella nucleases, was inhibited by the addition of EDTA. or storage at ‐20°C.