Termination of DNA synthesis by novel 3'-modifieddeoxyribonucleoside 5'-triphosphates (original) (raw)

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Department of Chemistry, Texas A & M University, College Station

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$ Present address: LI-COR Inc., Biotechnology Division, Lincoln, NE 685O4-500Or USA

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Department of Chemistry, Texas A & M University, College Station

TX 77843, USA

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1

Department of Chemistry, Texas A & M University, College Station

TX 77843, USA

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$ Present address: LI-COR Inc., Biotechnology Division, Lincoln, NE 685O4-500Or USA

Author Notes

Revision received:

25 July 1994

Published:

11 October 1994

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Michael L. Metzker, Ramesh Raghavachari, Stephen Richards, Swanee E. Jacutin, Andrew Civitello, Kevin Burgess, Richard A. Gibbs, Termination of DNA synthesis by novel 3'-modifieddeoxyribonucleoside 5'-triphosphates, Nucleic Acids Research, Volume 22, Issue 20, 11 October 1994, Pages 4259–4267, https://doi.org/10.1093/nar/22.20.4259
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Abstract

Eight 3′-modified-dNTPs were synthesized and tested in two different DNA template assays for incorporation activity. From this enzymatic screen, two 3′-O-methyldNTPs were shown to terminate DNA syntheses mediated by a number of polymerases and may be used as alternative terminators in Sanger sequencing. 3-O-(2-Nitrobenzyl)-dATP is a UV sensitive nucleotide and was shown to be incorporated by several thermostable DNA polymerases. Base specific termination and efficient photolytic removal of the 3′-protecting group was demonstrated. Following deprotection, DNA synthesis was reinitiated by the incorporation of natural nucleotides into DNA. The identification of this labile terminator and the demonstration of a one cycle stopstart DNA synthesis are initial steps in the development of a novel sequencing strategy.

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Author notes

$ Present address: LI-COR Inc., Biotechnology Division, Lincoln, NE 685O4-500Or USA

© 1994 Oxford University Press

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