Access

You are not currently logged in.

Access your personal account or get JSTOR access through your library or other institution:

login

Log in to your personal account or through your institution.

If You Use a Screen Reader

This content is available through Read Online (Free) program, which relies on page scans. Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.

NMR of Enzymatically Synthesized Uniformly 13C15N-Labeled DNA Oligonucleotides

Daniel P. Zimmer and Donald M. Crothers
Proceedings of the National Academy of Sciences of the United States of America
Vol. 92, No. 8 (Apr. 11, 1995), pp. 3091-3095
Stable URL: http://www.jstor.org/stable/2367010
Page Count: 5
  • Read Online (Free)
  • Subscribe ($19.50)
  • Cite this Item
Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.
NMR of Enzymatically Synthesized Uniformly 13C15N-Labeled DNA Oligonucleotides
Preview not available

Abstract

A procedure for the enzymatic synthesis of uniformly 13C15N-labeled DNA oligonucleotides in milligram quantities for NMR studies is described. Deoxynucleotides obtained from microorganisms grown on 13C and 15N nutrient sources are enzymatically phosphorylated to dNTPs, and the dNTPs are incorporated into oligonucleotides using a 3'-5' exonuclease-deficient mutant of Klenow fragment of DNA polymerase I and an oligonucleotide template primer designed for efficient separation of labeled product DNA from unlabeled template. The labeling strategy has been used to uniformly label one or the other oligonucleotide strand in the DNA duplex dGGCAAAACGG.dCCGTTTTGCC in order to facilitate assignment and structure determination by NMR. Application of 15N and 13C heteronuclear NMR experiments to isotopically labeled DNA is presented.

Page Thumbnails

  • Thumbnail: Page 
3091
    3091
  • Thumbnail: Page 
3092
    3092
  • Thumbnail: Page 
3093
    3093
  • Thumbnail: Page 
3094
    3094
  • Thumbnail: Page 
3095
    3095