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Published 2006 | public
Journal Article

Effects of a Modified Dye-Labeled Nucleotide Spacer Arm on Incorporation by Thermophilic DNA Polymerases


The ability of eight commercially available thermophilic DNA polymerases to sequentially incorporate fluorescently labeled nucleotides sequentially was analyzed by a gel based primer extension assay. Cy5-dUTP or a variant nucleotide in which the linker had been lengthened by 14 atoms between the dye and the nucleobase were compared. We found that the Cy5-dUTP with a longer linker resulted in longer primer extension lengths. Furthermore, some of the assayed polymerases are capable of extending the primer to the full or near full length of 30 nucleotides using dye-labeled nucleotides exclusively.

Additional Information

© 2006 Taylor & Francis Group, LLC. Received 19 February 2005; accepted 26 May 2005. This work was supported in part by DARPA, the National Institutes of Health, and the Rosen fellowship. The authors would like to thank Peter Dervan and his group for the use of their imager. C. L. thanks John DeModena and Saurabh Vyawahare for their useful advice and comments.

Additional details

August 19, 2023
October 24, 2023