title: Multiplexed single-molecule assay for enzymatic activity on flow-stretched DNA.
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.
authors: Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA.
authors: Sangjin
authors: S
authors: Kim
authors: Paul C
authors: PC
authors: Blainey
authors: Charles M
authors: CM
authors: Schroeder
authors: X Sunney
authors: XS
authors: Xie
mesh_terms: D017103:Bacillus Phages / Q000201:enzymology; D004247:DNA / Q000737:chemistry; D004277:DNA, Single-Stranded / Q000737:chemistry; D004279:DNA, Viral / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004548:Elasticity; D004926:Escherichia coli / Q000201:enzymology; D005090:Exodeoxyribonucleases / Q000378:metabolism*; D054303:HIV Reverse Transcriptase / Q000378:metabolism*; D008853:Microscopy / Q000379:methods; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D004277:DNA, Single-Stranded; D004279:DNA, Viral; D014764:Viral Proteins; D004247:DNA; D054303:HIV Reverse Transcriptase; D012321:DNA-Directed RNA Polymerases; D004259:DNA-Directed DNA Polymerase; D005090:Exodeoxyribonucleases; C043586:exo protein, Bacteriophage lambda