Nucleotide sequence analysis with polynucleotide kinase and nucleotide ‘mapping’ methods. 5′-Terminal sequence of deoxyribonucleic acid from bacteriophages λ and 424

Abstract

The polynucleotide kinase reaction was used in analyses of complex mixtures of oligodeoxynucleotides which were fractionated by various two-dimensional nucleotide ‘mapping’ procedures. Parallel ionophoretic analyses on DEAE-cellulose paper, pH2, and AE-cellulose paper, pH3.5, of venom phosphodiesterase partial digests of 5′-terminally labelled oligonucleotides enabled the sequence of the nucleotides to be deduced uniquely. A ‘diagonal ionophoresis’ method has been used with mixtures of nucleotides. Application of these methods to 5′-terminally labelled DNA from bacteriophage λ gave the terminal sequences pA-G-G-T-C-G and pG-G-G-C-G. Identical 5′-terminal sequences were found with DNA from bacteriophage 424.