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Nucleic Acids Research, 1977, Vol. 4, No. 10 3441-3454
© 1977

Articles

Use of specific endonuclease cleavage in RNA sequencing - an enzymic method for distinguishing between cytidine and uridine residues

Ramesh C. Gupta and Kurt Randerath

Department of Pharmacology, Baylor College of Medicine, Texas Medical Center, Houston TX 77030, USA

Received June 13, 1977. The extracellular ribonuclease I of the common slime mold Physarum polycephalum1 (RNase Phy1), which has recently been purified to homogeneity2, has been used to distinguish between C and U residues in 3'-end-labeled oligoribonucleotides. As shown by Bargetzi and coworkers2, 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 previous3 communication provide a direct means for identifying A, G, C, and U residues in end-labeled polyribonucleotides.


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A. Huttenhofer and J. Vogel
Experimental approaches to identify non-coding RNAs
Nucleic Acids Res., January 25, 2006; 34(2): 635 - 646.
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