Polymerase/DNA interactions and enzymatic activity: multi-parameter analysis with electro-switchable biosurfaces.

ABSTRACT: The engineering of high-performance enzymes for future sequencing and PCR technologies as well as the development of many anticancer drugs requires a detailed analysis of DNA/RNA synthesis processes. However, due to the complex molecular interplay involved, real-time methodologies have not been available to obtain comprehensive information on both binding parameters and enzymatic activities. Here we introduce a chip-based method to investigate polymerases and their interactions with nucleic acids, which employs an electrical actuation of DNA templates on microelectrodes. Two measurement modes track both the dynamics of the induced switching process and the DNA extension simultaneously to quantitate binding kinetics, dissociation constants and thermodynamic energies. The high sensitivity of

SUBMITTER: Langer A 

PROVIDER: S-EPMC4502528 | biostudies-literature | 2015 Jul

REPOSITORIES: biostudies-literature