Imaging of RNA in bacteria with self-ligating quenched probes.
We report on the application of a new class of oligonucleotide reporter probes, QUAL probes, that "light up" when a nucleophilic phosphorothioate probe binds adjacent to a dabsyl-quenched probe. These self-ligating DNA probes were used for sequence-specific detection of 16S rRNA in Escherichia coli cells. Strong fluorescence was observed only when the phosphorothioate and quenched dabsyl probes bind side-by-side on a 16S rRNA target. The results demonstrate the use of QUAL probes to detect specific RNA sequences in bacterial cells without enzymes and without washing steps.
Sando
Shinsuke
S
Department of Chemistry, Stanford University, Stanford, California 94305-5080, USA.
Kool
Eric T
ET
D015342:DNA Probes / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics*; D005456:Fluorescent Dyes / Q000737:chemistry*; D012329:RNA, Bacterial / Q000032:analysis* / Q000737:chemistry; D012336:RNA, Ribosomal, 16S / Q000032:analysis* / Q000737:chemistry
D015342:DNA Probes; D005456:Fluorescent Dyes; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S