Screening for ribosomal-based false positives following prokaryotic mRNA differential display.
Differential display (DD) and the closely related RNA arbitrarily primed PCR (RAP-PCR) have become the molecular tools of choice for identifying and isolating differentially expressed genes in both eukaryotic and prokaryotic systems. However, one of the current drawbacks of both techniques is the high number of false positives generated. In prokaryotic applications, the many false positive typically generated by DD are subsequently identified as rRNAs because of their greater abundance compared to mRNAs. To circumvent this problem, full-length 16S and 23S rDNA probes, derived from Pseudomonas putida G7 and Pseudomonas aeruginosa FRD1, respectively, were used as a prescreening approach to discriminate between those bands, which appear to be differentially expressed mRNAs, but in fact are rRNAs, following prokaryotic mRNA DD.
Nagel
A C
AC
University of Tennessee Knoxville, USA
Fleming
J T
JT
Sayler
G S
GS
Beattie
K L
KL
D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D018076:DNA, Complementary / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005189:False Positive Reactions; D016133:Polymerase Chain Reaction / Q000379:methods; D011550:Pseudomonas aeruginosa / Q000235:genetics; D016958:Pseudomonas putida / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis*; D012335:RNA, Ribosomal / Q000032:analysis*; D012336:RNA, Ribosomal, 16S / Q000235:genetics / Q000302:isolation & purification; D012338:RNA, Ribosomal, 23S / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
D015342:DNA Probes; D004269:DNA, Bacterial; D018076:DNA, Complementary; D012333:RNA, Messenger; D012335:RNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S