Stackebrandt E, Ludwig W, Schleifer K H, Gross H J
J Mol Evol. 1981;17(4):227-36. doi: 10.1007/BF01732760.
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.
本文介绍了一种用于系统发育研究的快速、简单的寡核苷酸编目方法。该方法包括对核糖体16S RNA的核糖核酸酶T1抗性寡核苷酸进行体外5'-32P标记,以及通过高压电泳和梯度薄层色谱进行指纹分析。寡核苷酸序列通过迁移率变动法确定,使用可控碱裂解、高压电泳和同系色谱法。这些程序特别有助于分析长的核糖核酸酶T1抗性寡核苷酸。建立了三种放线菌的寡核苷酸编目,即土栖链霉菌、菲律宾游动放线菌和规则瓶孢放线菌。这些编目与通过桑格和沃斯描述的方法获得的编目相当。