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. 1965 Feb;94(2):314–322. doi: 10.1042/bj0940314

Methods of automatic nucleotide-sequence analysis. Multicomponent spectrophotometric analysis of mixtures of nucleic acid components by a least-squares procedure

Sheila Lee 1,2, D McMullen 1,2, G L Brown 1,2, A R Stokes 1,2
PMCID: PMC1206512  PMID: 14346087

Abstract

1. A theoretical analysis of the errors in multicomponent spectrophotometric analysis of nucleoside mixtures, by a least-squares procedure, has been made to obtain an expression for the error coefficient, relating the error in calculated concentration to the error in extinction measurements. 2. The error coefficients, which depend only on the `library' of spectra used to fit the experimental curves, have been computed for a number of `libraries' containing the following nucleosides found in s-RNA: adenosine, guanosine, cytidine, uridine, 5-ribosyluracil, 7-methylguanosine, 6-dimethylaminopurine riboside, 6-methylaminopurine riboside and thymine riboside. 3. The error coefficients have been used to determine the best conditions for maximum accuracy in the determination of the compositions of nucleoside mixtures. 4. Experimental determinations of the compositions of nucleoside mixtures have been made and the errors found to be consistent with those predicted by the theoretical analysis. 5. It has been demonstrated that, with certain precautions, the multicomponent spectrophotometric method described is suitable as a basis for automatic nucleotide-composition analysis of oligonucleotides containing nine nucleotides. Used in conjunction with continuous chromatography and flow chemical techniques, this method can be applied to the study of the sequence of s-RNA.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. APGAR J., HOLLEY R. W., MERRILL S. H. Purification of the alanine-, valine-, histidine-, and tyrosine-acceptor ribonucleic acids from yeast. J Biol Chem. 1962 Mar;237:796–802. [PubMed] [Google Scholar]
  2. COHN W. E. 5-Ribosyl uracil, a carbon-carbon ribofuranosyl nucleoside in ribonucleic acids. Biochim Biophys Acta. 1959 Apr;32:569–571. doi: 10.1016/0006-3002(59)90644-4. [DOI] [PubMed] [Google Scholar]
  3. COHN W. E. Pseudouridine, a carbon-carbon linked ribonucleoside in ribonucleic acids: isolation, structure, and chemical characteristics. J Biol Chem. 1960 May;235:1488–1498. [PubMed] [Google Scholar]
  4. DAVIS F. F., ALLEN F. W. Ribonucleic acids from yeast which contain a fifth nucleotide. J Biol Chem. 1957 Aug;227(2):907–915. [PubMed] [Google Scholar]
  5. DEKKER C. A. Nucleic acids. Selected topics related to their enzymology and chemistry. Annu Rev Biochem. 1960;29:453–474. doi: 10.1146/annurev.bi.29.070160.002321. [DOI] [PubMed] [Google Scholar]
  6. FLEISSNER E., BOREK E. A new enzyme of RNA synthesis: RNA methylase. Proc Natl Acad Sci U S A. 1962 Jul 15;48:1199–1203. doi: 10.1073/pnas.48.7.1199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. FOX J. J., SHUGAR D. Spectrophotometric studies of nucleic acid derivatives and related compounds as a function of pH. II. Natural and synthetic pyrimidine nucleosides. Biochim Biophys Acta. 1952 Oct;9(4):369–384. doi: 10.1016/0006-3002(52)90181-9. [DOI] [PubMed] [Google Scholar]
  8. GOLDSTEIN J., BENNETT T. B., CRAIG L. C. COUNTERCURRENT DISTRIBUTION STUDIES OF E. COLI B SRNA. Proc Natl Acad Sci U S A. 1964 Jan;51:119–125. doi: 10.1073/pnas.51.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. LAWLEY P. D., BROOKES P. FURTHER STUDIES ON THE ALKYLATION OF NUCLEIC ACIDS AND THEIR CONSTITUENT NUCLEOTIDES. Biochem J. 1963 Oct;89:127–138. doi: 10.1042/bj0890127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LITTLEFIELD J. W., DUNN D. B. The occurrence and distribution of thymine and three methylated-adenine bases in ribonucleic acids from several sources. Biochem J. 1958 Dec;70(4):642–651. doi: 10.1042/bj0700642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. MICHELSON A. M., COHN W. E. Cyclo-pseudouridine and the configuration of pseudouridine. Biochemistry. 1962 May 25;1:490–495. doi: 10.1021/bi00909a020. [DOI] [PubMed] [Google Scholar]
  12. REID J. C., PRATT A. W. Vector analysis of ultraviolet mixture spectra: the composition of ribonucleic acid. Biochem Biophys Res Commun. 1960 Oct;3:337–342. doi: 10.1016/0006-291x(60)90040-1. [DOI] [PubMed] [Google Scholar]
  13. SKEGGS L. T., Jr An automatic method for colorimetric analysis. Am J Clin Pathol. 1957 Sep;28(3):311–322. doi: 10.1093/ajcp/28.3_ts.311. [DOI] [PubMed] [Google Scholar]
  14. SMITH J. D., DUNN D. B. The occurrence of methylated guanines in ribonucleic acids from several sources. Biochem J. 1959 Jun;72(2):294–301. doi: 10.1042/bj0720294. [DOI] [PMC free article] [PubMed] [Google Scholar]

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