Use of the SYBR Green dye for measuring helicase activity

Sammer Siddiqui, Irfan Khan, Shamshad Zarina, Syed Ali

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Here we describe a non-radioactive assay that exploits the fluorescent dye SYBR Green to measure the helicase enzyme activity. SYBR Green I emits fluorescence upon intercalation with double-stranded DNA or RNA. The fluorescence is lost proportionally as the nucleic acid is converted to single strands by a helicase, and this decrease in fluorescence intensity can be used to measure the activity of the helicase enzyme. The reaction was prepared by mixing a double-stranded substrate with the helicase enzyme, buffer, ATP and SYBR Green I. After completion, the reaction was terminated by EDTA and fluorescence was measured. Using this technique, a linear increase in substrate release was observed with increasing time and helicase concentrations. The assay described here is speedy, efficient and economical; it holds promise for use in large-scale screening of drugs that target helicases.

Original languageEnglish
Pages (from-to)196-198
Number of pages3
JournalEnzyme and Microbial Technology
Volume52
Issue number3
DOIs
Publication statusPublished - Mar 5 2013
Externally publishedYes

Fingerprint

Coloring Agents
Dyes
Fluorescence
Assays
Enzymes
Preclinical Drug Evaluations
Double-Stranded RNA
Adenosinetriphosphate
Nucleic acids
Ethylenediaminetetraacetic acid
Enzyme activity
Substrates
Intercalation
RNA
Fluorescent Dyes
Edetic Acid
Nucleic Acids
Buffers
Screening
DNA

Keywords

  • Helicase
  • SYBR Green

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Use of the SYBR Green dye for measuring helicase activity. / Siddiqui, Sammer; Khan, Irfan; Zarina, Shamshad; Ali, Syed.

In: Enzyme and Microbial Technology, Vol. 52, No. 3, 05.03.2013, p. 196-198.

Research output: Contribution to journalArticle

Siddiqui, Sammer ; Khan, Irfan ; Zarina, Shamshad ; Ali, Syed. / Use of the SYBR Green dye for measuring helicase activity. In: Enzyme and Microbial Technology. 2013 ; Vol. 52, No. 3. pp. 196-198.
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