Rapid cataloging of ribonuclease T1 resistant oligonucleotides from ribosomal RNAs for phylogenetic studies

Abstract

A rapid and simple method of oligonucleotide cataloging for phylogenetic studies is presented. It involves in vitro 5′-³²P-labelling of RNase T₁ - resistant oligonucleotides of ribosomal 16S RNA and fingerprinting by high voltage electrophoresis and gradient thinlayer chromatography. Oligonucleotide sequences are established by the mobility shift method, using controlled alkali cleavage, high voltage electrophoresis and homochromatography. These procedures facilitate in particular the analysis of long RNase T₁ - resistant oligonucleotides. Oligonucleotide catalogs are established for three Actinomycetes, namelyOerskovia turbata, Actinoplanes philippinensis andAmpullariella regularis. These catalogs are equivalent to those obtained by methods which were described by Sanger and Woese.