Detection of bacterial mRNA using polymerase chain reaction.
A method was developed for the detection of bacterial mRNAs using reverse transcriptase followed by the polymerase chain reaction (PCR) and Southern blot analysis. The method involves brief inhibition of protein synthesis with chloramphenicol, followed by reverse transcription, PCR amplification of cDNA and Southern blot hybridization. Detection of mRNAs by reverse transcription-PCR-Southern blot analysis is orders of magnitude more sensitive than Northern blot hybridization.
Mahbubani
M H
MH
Dept. of Biology, University of Louisville, KY 40292.
Bej
A K
AK
Miller
R D
RD
Atlas
R M
RM
DiCesare
J L
JL
Haff
L A
LA
D015139:Blotting, Southern; D002701:Chloramphenicol / Q000494:pharmacology; D007875:Legionella / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000032:analysis*; D012333:RNA, Messenger / Q000032:analysis; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism
D012329:RNA, Bacterial; D012333:RNA, Messenger; D002701:Chloramphenicol; D012194:RNA-Directed DNA Polymerase