Section 37
Chapter 36,709

Use of the SYBR Green dye for measuring helicase activity

Siddiqui, S.; Khan, I.; Zarina, S.; Ali, S.

Enzyme and Microbial Technology 52(3): 196-198


ISSN/ISBN: 1879-0909
PMID: 23410932
DOI: 10.1016/j.enzmictec.2012.12.008
Accession: 036708082

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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.

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