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PMID: 5119768 Published · ppublish English Journal Article

Chromatographic separation of mononucleotides derived from transfer ribonucleic acids.

The Biochemical journal ·Vol. 121 ·No. 3 ·1971-02-00 ·Pages 371-7

Holness NJ, Atfield G

Abstract

1. A method is described for fractionating alkaline and enzymic hydrolysates of ribonucleic acids and oligonucleotides on DEAE-cellulose columns by using a constant-composition eluent. 2. The relative partition coefficients of the four major and several minor nucleotides present in nucleic acid digests are given. 3. The influence of pH and of molarity of the buffer on the separation is described. 4. Relations for the quantitative determination of the major nucleotides from their absorption zone areas on the chromatographic record are derived. 5. Hydrolysis of oligonucleotides with T2 ribonuclease yields solely the nucleoside 3'-monophosphates, thereby simplifying the subsequent chromatographic separation. 6. Examples of the qualitative and quantitative analysis of oligonucleotides from enzymic hydrolysates of RNA are given.

MeSH Terms
Chromatography, DEAE-Cellulose Hydrogen-Ion Concentration Hydrolysis Methods Molecular Weight Nucleotides/isolation & purification Polynucleotides/isolation & purification RNA/isolation & purification RNA, Transfer/analysis Ribonucleases
Chemicals
Nucleotides Polynucleotides RNA RNA, Transfer Ribonucleases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Holness N J
Atfield G
References (8)
8 references, click to expand
  1. Ion-exchange chromatography of nucleotides on polyethyleneimine cellulose columns: analysis of maize grain extracts.
    Anal Biochem. 1968 Jan;22(1):35-46 PMID: 5636956
  2. Rapid ion-exchange chromatographic microanalysis of ultraviolet-absorbing materials and its application to nucleosides.
    Anal Biochem. 1968 Oct 24;25(1):77-98 PMID: 5704775
  3. Column chromatography of phosphorylated nucleosides and deoxynucleosides on PEI-cellulose.
    Anal Biochem. 1969 Feb;27(2):306-10 PMID: 5767186
  4. Analyses of pancreatic ribonuclease digests of Torulopsis utilis transfer ribonucleic acid's specific for alanine, valine and several other amino acids.
    J Biochem. 1969 Feb;65(2):159-69 PMID: 5814320
  5. Rapid analysis of ribonucleosides and bases at the picomole level using pellicular cation exchange resin in narrow bore columns.
    Anal Chem. 1969 Aug;41(10):1227-34 PMID: 5797315
  6. Investigations into the nucleotide sequence of cysteine transfer ribonucleic acid from baker's yeast.
    Biochem J. 1971 Jan;121(1):26P PMID: 5116547
  7. The separation of biochemically important substances by ion exchange chromatography.
    Ann N Y Acad Sci. 1953 Nov 11;57(3):204-24 PMID: 13139317
  8. A NEW METHOD FOR THE DETERMINATION OF THE BASE COMPOSITION OF RIBONUCLEIC ACID.
    J Biol Chem. 1963 Sep;238:3065-7 PMID: 14081925
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
Published
1971-02-00
Pages
371-7
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1176583
Subset
IM
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