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High resolution chromatography of ribonucleosides and its application to RNA analysis
Author(s) -
Tanaka Kouji,
Yazawa Kouhei,
Nakajima Terumi
Publication year - 1989
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130030604
Subject(s) - rnase p , oligonucleotide , chemistry , nucleoside , transfer rna , chromatography , rna , rnase h , biochemistry , dna , gene
A simple and precise method was developed for the separation of nucleosides including modified nucleosides and oligonucleotides. Nineteen kinds of nucleosides were completely separated by HPLC using an ODS column (TSK‐gel ODS 80TM) and aqueous mobile phases. The RNA molecule was digested by base restrictive RNase (RNase A, RNase T 1 ) and the digests were separated chromatographically into each oligonucleotide. The nucleoside composition of an oligonucleotide was then determined by this analytical system. It is thus possible to fit the oligonucleotide in the original RNA molecule by using modified bases as markers. The reaction site of quinacrine mustard for tRNA Phe (from yeast) could be determined by this analytical system.