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

Sangjin Kim, Paul C. Blainey, Charles M. Schroeder, X. Sunney Xie

Research output: Contribution to journalArticle

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 ∼10 nm mechanical stability over the timescale of hours. With multiplexed data collection, we applied the assay to φ29 DNA polymerase, HIV-1 reverse transcriptase, λ exonuclease and Escherichia coli RNA polymerase.

Original languageEnglish (US)
Pages (from-to)397-399
Number of pages3
JournalNature Methods
Volume4
Issue number5
DOIs
StatePublished - May 1 2007
Externally publishedYes

Fingerprint

Exonucleases
Enzyme Assays
DNA-Directed DNA Polymerase
DNA-Directed RNA Polymerases
Nucleic Acids
Assays
Escherichia coli
Molecules
DNA
Enzymes
Mechanical stability
Human immunodeficiency virus 1 reverse transcriptase

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Multiplexed single-molecule assay for enzymatic activity on flow-stretched DNA. / Kim, Sangjin; Blainey, Paul C.; Schroeder, Charles M.; Xie, X. Sunney.

In: Nature Methods, Vol. 4, No. 5, 01.05.2007, p. 397-399.

Research output: Contribution to journalArticle

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