Search National Agricultural Library Digital Collections
Back to Search
NALDC Record Details:
DNA extraction protocol for rapid PCR detection of pathogenic bacteria
Twelve reagents were evaluated to develop a direct DNA extraction method suitable for PCR detection of foodborne bacterial pathogens. Many reagents exhibited strong PCR inhibition, requiring significant dilution of the extract with a corresponding reduction in sensitivity. Most reagents also exhibited much lower recovery of DNA from the gram-positive test organism (Listeria monocytogenes) than from the gram-negative organism (Escherichia coli O157:H7), preventing unbiased detection and quantitation of both organisms. The 5 HotSHOT + Tween reagent exhibited minimal inhibition and high extraction efficiency for both test organisms, providing a 15-min single-tube DNA-extraction protocol suitable for highly sensitive quantitative PCR assays.
Jeffrey D. Brewster
George C. Paoli
Escherichia coli O157
animal pathogenic bacteria
polymerase chain reaction
Analytical Biochemistry 2013 7 16 v.442 no.1
Journal Articles, USDA Authors, Peer-Reviewed
Download [PDF File]
Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted.
Agricultural Research Service
Web Policies and Important Links