Use of specific endonuclease cleavage in RNA sequencing - an enzymic method for distinguishing between cytidine and uridine residues
- 1 January 1977
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 4 (10), 3441-3454
- https://doi.org/10.1093/nar/4.10.3441
Abstract
The extracellular ribonuclease I of the common slime mold physarum polycephalum (RNase Phy1), which has recently been purified to homogeneity, has been used to distinguish between C and U residues in 3'-end-labeled oligoribonucleotides. As shown by Bargetzi and coworkers, this enzyme exhibits strong cleavage preference for U-N over C-N and N-C over N-U bonds. In the present paper, conditions are being detailed, which enable one to deduce the sequences of rather large, pyrimidine-rich, terminally labeled oligonucleotides by partial digestion with RNases U2, A, and Phy1, followed by resolution of the cleavage products by size. The techniques described in this and a previous communication provide a direct means for identifying A, G, C, and U residues in end-labeled polyribonucleotides.Keywords
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