APPROACHES TO SEQUENCE-ANALYSIS OF I-125-LABELED RNA

A method is described for the initial steps of sequence analysis of RNase Tl- and pancreatic RNase-resistant oligonucleotides of RNA containing cytidylate residues labeled in vitro with 125I. in many cases an oligo-nucleotide sequence can be deduced from a consideration of (i) its relative position in the two-dimensional fingerprint (with DEAE thin layer homo-chromatographic second dimension), (ii) its electrophoretic mobility on DEAE paper at pH 1.9, and (iii) identification of its products of further enzymatic digestion by comparison with a set of marker oligonucleotides. Additional methods including analysis of oligonucleotides following chemical blocking of uridylate residues with CMCT and analysis of products of incomplete enzymatic digestion are also discussed. © 1979 Information Retrieval Limited.