Efficiency of the polymerase chain reaction amplification of the uid gene for detection of Escherichia coli in contaminated water.
Direct detection of Escherichia coli from polluted river water was achieved using polymerase chain reaction (PCR) amplification of the uid gene. Amplification using DNA from environmental samples resulted in non-specific DNA fragments. Specific amplification was achieved through use of the touch-down PCR procedure. Targeting the uidA structural region of the gene gave reproducibly better amplification than targeting the uidR regulatory region. The data demonstrate conditions for optimal specific detection.
Iqbal
S
S
Department of Genetics and Microbiology, University of Liverpool, UK.
Robinson
J
J
Deere
D
D
Saunders
J R
JR
Edwards
C
C
Porter
J
J
D001431:Bacteriological Techniques; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification*; D005069:Evaluation Studies as Topic; D005798:Genes, Bacterial; D005809:Genes, Regulator; D005966:Glucuronidase / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012722:Sewage / Q000382:microbiology; D014871:Water Microbiology
D004269:DNA, Bacterial; D012722:Sewage; D005966:Glucuronidase