PCR-based preparation of 23S rRNA-targeted group-specific polynucleotide probes

• DOI: 10.1128/aem.60.9.3236-3244.1994
• Publication Date: 2020-01-01T04:59:58Z
• Authors: W Ludwig; S Dorn; N Springer; G Kirchhof; K H Schleifer
• Keywords: Hybridization probe; 23S ribosomal RNA; Molecular probe; Acinetobacter calcoaceticus

DNA coding for a variable region within domain III of bacterial 23S rRNA was used as the target group-specific polynucleotide hybridization probes. The corresponding rDNA amplified in vitro by PCR technique combination with pair primers specific flanking conserved sites. fragments were cloned or directly RNA probes generated transcription rDNA. labeled incorporating modified nucleotides during amplification random priming. use transcribed single-stranded instead double-stranded provided stronger signals. Group-specific prepared from genomic DNAs cells Acinetobacter calcoaceticus, Alcaligenes faecalis, Aeromonas hydrophila, Nannocystis exedens, Pseudomonas aeruginosa, fluorescens, and stutzeri.