Home LiteratureArticle Details
PMID: 331267 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Use of specific endonuclease cleavage in RNA sequencing.

Nucleic acids research ·Vol. 4 ·No. 6 ·1977-06-00 ·Pages 1957-78

Gupta RC, Randerath K

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.

MeSH Terms
Base Sequence Endonucleases/metabolism Escherichia coli Methods Oligoribonucleotides RNA, Transfer Ribonucleases Yeasts
Chemicals
Oligoribonucleotides RNA, Transfer Endonucleases Ribonucleases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Gupta R C
Randerath K
References (30)
30 references, click to expand
  1. THE EFFECT OF UREA, FORMAMIDE, AND GLYCOLS ON THE SECONDARY BINDING FORCES IN THE ION-EXCHANGE CHROMATOGRAPHY OF POLYNUCLEOTIDES OF DEAE-CELLULOSE.
    Biochemistry. 1963 Jul-Aug;2:697-702 PMID: 14075100
  2. Nucleotide sequences at the 5'termini of rabbit alpha and beta globin mRNA.
    Cell. 1976 Dec;9(4 PT 2):747-60 PMID: 189943
  3. Nucleotide sequences from the rabbit beta globin gene inserted into Escherichia coli plasmids.
    Science. 1977 Jan 28;195(4276):389-91 PMID: 318762
  4. A new method for sequencing DNA.
    Proc Natl Acad Sci U S A. 1977 Feb;74(2):560-4 PMID: 265521
  5. The first nucleotide sequence of an organelle transfer RNA: chloroplastic tRNAphe.
    Cell. 1976 Dec;9(4 PT 2):717-23 PMID: 828079
  6. A double-labeling procedure for sequence analysis of picomole amounts of nonradioactive RNA fragments.
    Nucleic Acids Res. 1976 Nov;3(11):2895-914 PMID: 826884
  7. Sequence analysis of small amounts of nonradioactive oligoribonucleotides containing ribose-methylated nucleosides by a combination of 3H- and 32P-labeling techniques.
    Anal Biochem. 1976 Nov;76(l):269-80 PMID: 793443
  8. Nucleotide sequence of Escherichia coli tyrosine transfer ribonucleic acid.
    Science. 1969 Feb 14;163(3868):693-5 PMID: 4883890
  9. Studies on polynucleotides. LXXXII. Yeast phenylalanine transfer ribonucleic acid: partial digestion with ribonuclease T-1 and derivation of the total primary structure.
    J Biol Chem. 1968 Feb 10;243(3):598-608 PMID: 5637712
  10. Studies on extracellular ribonucleases of Ustilago sphaerogena. Characterization of substrate specificity with special reference to purine-specific ribonucleases.
    Biochem J. 1968 Feb;106(3):609-13 PMID: 5639916
  11. A ribonuclease from Physarum. Biochemical properties and synthesis in the mitotic cycle.
    Biochim Biophys Acta. 1969 Nov 19;195(1):87-98 PMID: 5390539
  12. Studies on ribonucleases from Physarum polycephalum. Purification and characterization of substrate specificity.
    J Biochem. 1969 May;65(5):693-700 PMID: 5817396
  13. Sequence analysis of nonradioactive RNA fragments by periodate-phosphatase digestion and chemical tritium labeling: characterization of large oligonucleotides and oligonucleotides containing modified nucleosides.
    Nucleic Acids Res. 1974 Sep;1(9):1121-41 PMID: 4375807
  14. Base analysis of ribopolynucleotides by chemical tritium labeling: a methodological study with model nucleosides and purified tRNA species.
    Anal Biochem. 1972 Jul;48(1):172-98 PMID: 4339367
  15. An evaluation of film detection methods for weak beta-emitters, particularly tritium.
    Anal Biochem. 1970 Mar;34:188-205 PMID: 4314971
  16. The nucleotide sequences of tyrosine transfer RNAs of Escherichia coli.
    Eur J Biochem. 1970 Apr;13(3):461-83 PMID: 4315419
  17. Absence of the sequence G-T-psi-C-G(A)- in several eukaryotic cytoplasmic initiator transfer RNAs.
    Proc Natl Acad Sci U S A. 1973 Apr;70(4):1041-5 PMID: 4577787
  18. The corrected nucleotide sequence of yeast leucine transfer ribonucleic acid.
    Biochem Biophys Res Commun. 1973 Apr 16;51(4):951-5 PMID: 4574158
  19. The effects of polyamines on a residue-specific human plasma ribonuclease.
    J Biol Chem. 1975 Mar 25;250(6):2206-12 PMID: 234961
  20. Chromatography of 32P-labelled oligonucleotides on thin layers of DEAE-cellulose.
    Eur J Biochem. 1969 Dec;11(2):395-9 PMID: 4902612
  21. Nucleotide sequence of the "denaturable" leucine transfer RNA from yeast.
    Science. 1971 Apr 23;172(3981):385-7 PMID: 4927676
  22. A method for the isolation of cytidylate series from ribonuclease T1-oligonucleotides.
    Anal Biochem. 1975 Jul;67(1):319-26 PMID: 807124
  23. A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase.
    J Mol Biol. 1975 May 25;94(3):441-8 PMID: 1100841
  24. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography.
    Eur J Biochem. 1975 Aug 15;56(2):335-41 PMID: 1175627
  25. Use of polynucleotide kinase in fingerprinting non-radioactive nucleic acids.
    J Mol Biol. 1969 Aug 14;43(3):607-17 PMID: 5401237
  26. Studies on polynucleotides. LXXIX. Yeast phenylalanine transfer ribonucleic acid: products obtained by degradation with pancreatic ribonuclease.
    J Biol Chem. 1968 Feb 10;243(3):575-83 PMID: 5637709
  27. Studies on polynucleotides. LXXVII. The labeling of end groups in polynucleotide chains: the selective modification of diol end groups in ribonucleic acids.
    J Biol Chem. 1968 Feb 10;243(3):556-64 PMID: 5637707
  28. Ion-exchange thin-layer chromatography. XV. Preparation, properties and applications of paper-like PEI-cellulose sheets.
    J Chromatogr. 1966 Apr;22(1):110-7 PMID: 5922282
  29. Tritium sequence analysis of oligoribonucleotides: a combination of post-labeling and thin-layer chromatographic techniques for the analysis of partial snake venom phosphodiesterase digests.
    Nucleic Acids Res. 1974 Oct;1(10):1329-41 PMID: 10793693
  30. Fingerprinting nonradioactive ribonucleic acid with the aid of polynucleotide phosphorylase.
    Nucleic Acids Res. 1974 Jan;1(1):171-81 PMID: 10793669
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/4.6.1957
Published
1977-06-00
Pages
1957-78
Language
English
Region
England
NLM ID
0411011
PMCID
PMC342534
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/4.6.1957 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2