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Rapid cataloging of ribonuclease T1 resistant oligonucleotides from ribosomal RNAs for phylogenetic studies
Authors:Erko Stackebrandt  Wolfgang Ludwig  Karl-Heinz Schleifer  Hans J. Gross
Affiliation:1. Department of Microbiology, Technische Universit?t München, Arcisstr. 21, D-8000, München 2, Germany
2. Max-Planck-Institut für Biochemie, D-8033, Martinsried bei München, Germany
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 finger-printing 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 fo three Actinomycetes, namely Oerskovia turbata, Actinoplanes philippinensis and Ampullariella regularis. These catalogs are equivalent to those obtained by methods which were described by Sanger and Woese.
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