Sequence specific effects on the incorporation of dideoxynucleotides by a modified T7 polymerase

Citation

Blanchard, Alan-Philippe (1993) Sequence specific effects on the incorporation of dideoxynucleotides by a modified T7 polymerase. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/9vz4-0e68. https://resolver.caltech.edu/CaltechTHESIS:11122012-105407501

Abstract

While incorporating nucleotides onto the end of a DNA molecule, DNA polymerases selectively discriminate against dideoxynucleotides in favor of incorporating deoxynucleotides. The magnitude of this discrimination is modulated by the template DNA sequence near the incorporation site. This effect has been characterized by analyzing the raw data from a large number of DNA sequencing experiments. It is shown that, for bacteriophage T7 polymerase, the 5 contiguous bases extending from 3 bases 3' (on the template strand) from the incorporation site to 1 base 5' of the incorporation site are the most important in modulating dideoxynucelotide discrimination. A table of discrimination ratios for 1007 different 5-mer contexts is presented.

Thesis Files