Mass-spectrometry analysis of modifications at DNA termini induced by DNA polymerases.

ABSTRACT: Non-natural nucleotide substrates are widely used in the enzymatic synthesis of modified DNA. The terminal activity of polymerases in the presence of modified nucleotides is an important, but poorly characterized, aspect of enzymatic DNA synthesis. Here, we studied different types of polymerase activity at sequence ends using extendable and non-extendable synthetic models in the presence of the Cy5-dUTP analog Y. In primer extension reactions with selected exonuclease-deficient polymerases, nucleotide Y appeared to be a preferential substrate for non-templated 3'-tailing, as determined by MALDI mass-spectrometry and gel-electrophoresis. This result was further confirmed by the 3'-tailing of a non-extendable hairpin oligonucleotide model. Additionally, DNA polymerases induce an exchange of

SUBMITTER: Smirnov IP 

PROVIDER: S-EPMC5532294 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature