A different approach to RNA sequencing

Abstract

DURING the past few years techniques to simplify the systematic sequence determination of nucleic acids have become available; these involve the use of polyacrylamide gels from which the sequence may be directly and rapidly read off. The most recent of these, applicable to RNA fragments, rely on the base-specific nature of cleavage by T₁ and U₂ RNases^1,2 and pancreatic RNase and RNase I from Physarum polycephalum² by which a spectrum of 5′ terminally-labelled fragments may be produced and comigrated with a ‘ladder’ achieved by incubating the fragment with alkali or hot formamide. We describe here a method for the sequence determination of RNA fragments bearing a blocked 5′ terminus, in which the above enzymatic hydrolysis steps and their accompanying problems may be avoided.