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PMID: 202925 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The use of nuclease P1 in sequence analysis of end group labeled RNA.

Nucleic acids research ·Vol. 4 ·No. 12 ·1977-12-00 ·Pages 4091-108

Silberklang M, Gillum AM, RajBhandary UL

Abstract

A method is described for the direct sequence analysis of 20-25 nucleotides from the termini of 5'- or 3'-end-group [32P] labeled RNA. The method involves partial endonucleolytic digestion of the labeled RNA with nuclease P1 (from Penicillium citrinum) followed by separation of the partial digestion products by two-dimensional homochromatography, the nucleotide sequence being determined by mobility shift analysis. This procedure has been applied to the sequence analysis of the terminal regions of tRNAs and of high molecular weight RNA, such as messenger RNA or viral RNA. A further application involves its use in conjunction with snake venom phosphodiesterase to determine the sequence of 5'-end group labeled oligonucleotides, containing modified bases, derived from T1 or pancreatic RNase digestion of tRNA.

MeSH Terms
Base Sequence Isotope Labeling Oligoribonucleotides/analysis Phosphoric Diester Hydrolases Phosphorus Radioisotopes RNA, Transfer Ribonucleases
Chemicals
Oligoribonucleotides Phosphorus Radioisotopes RNA, Transfer Ribonucleases Phosphoric Diester Hydrolases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Silberklang M
Gillum A M
RajBhandary U L
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22 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/4.12.4091
Published
1977-12-00
Pages
4091-108
Language
English
Region
England
NLM ID
0411011
PMCID
PMC343228
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/4.12.4091 PMC Full Text View journal details
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