PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of narLink to Publisher's site
 
Nucleic Acids Res. Oct 11, 1994; 22(20): 4259–4267.
PMCID: PMC331938
Termination of DNA synthesis by novel 3'-modified-deoxyribonucleoside 5'-triphosphates.
M L Metzker, R Raghavachari, S Richards, S E Jacutin, A Civitello, K Burgess, and R A Gibbs
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030.
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-methyl-dNTPs 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 stop-start DNA synthesis are initial steps in the development of a novel sequencing strategy.
Full text
Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.2M), or click on a page image below to browse page by page.
Images in this article
Click on the image to see a larger version.
Articles from Nucleic Acids Research are provided here courtesy of
Oxford University Press