Multiplexed single-molecule assay for enzymatic activity on flow-stretched DNA.

Kim, Sangjin; Blainey, Paul C; Schroeder, Charles M; Xie, X Sunney · Nat Methods · 2007

basic_science · Level V

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Abstract

We report a single-molecule assay for nucleic-acid enzymes on flow-stretched DNA templates. To facilitate the detection of slow or intermittent enzymatic activities, we developed the assay with 15-nm spatial resolution at a frame rate of 1 Hz and approximately 10 nm mechanical stability over the timescale of hours. With multiplexed data collection, we applied the assay to phi29 DNA polymerase, HIV-1 reverse transcriptase, lambda exonuclease and Escherichia coli RNA polymerase.

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