PCR products generated from unpurified Salmonella DNA are degraded by thermostable nuclease activity

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 pr...

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Bibliographic Details
Published inLetters in applied microbiology Vol. 16; no. 2; pp. 59 - 61
Main Authors Gibson, J.R, McKee, R.A
Format Journal Article
LanguageEnglish
Published England 01.02.1993
Subjects
Base Sequence
Deoxyribonucleases - metabolism
DNA
DNA, Bacterial - genetics
DNA, Bacterial - metabolism
Enzyme Stability
food contamination
heat stability
hydrolysis
microbial contamination
Molecular Sequence Data
nucleases
Polymerase Chain Reaction
Salmonella
Salmonella - genetics
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Summary: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 degrees C. This post-PCR DNA degradation, attributed to endogenous Salmonella nucleases, was inhibited by the addition of EDTA, or storage at -20 degrees C.
ISSN:0266-8254
1472-765X
DOI:10.1111/j.1472-765X.1993.tb00342.x