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. 1977 Jun;4(6):1957-78.
doi: 10.1093/nar/4.6.1957.

Use of specific endonuclease cleavage in RNA sequencing

R C Gupta, K Randerath
Free PMC article

Use of specific endonuclease cleavage in RNA sequencing

R C Gupta et al. Nucleic Acids Res. 1977 Jun.
Free PMC article

Abstract

Nonradioactive RNA fragments may be sequenced by incorporation of (3H)-label into 3'-terminal positions, controlled digestion with specific ribonucleases, and separation according to size of the digestion products on polyethyleneimine- (PEI-) cellulose thin layers. This combination of techniques allows one to measure accurately distances of specific cleavage sites from the labeled terminal positions. The cleavage specificities of RNases T1, U2, and A are utilized to identify the positions of G, A, and pyrimidine residues respectively. C and U may be distinguished by mobility differences on PEI-cellulose thin layers at ph 2.6. The procedure is simple, rapid, and highly sensitive; as little as 0.5 - 1 microgram of a RNA of the size of tRNA will be needed to sequence all fragments in a complete RNase digest.

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References

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