Use of the SYBR Green dye for measuring helicase activity.

Sammer Siddiqui, Irfan Khan, Shamshad Zarina, Syed Ali

Department of Biological and Biomedical Sciences, The Aga Khan University, Karachi, Pakistan.

Enzyme and microbial technology 2013 Mar 5

filter terms:

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. Copyright © 2013 Elsevier Inc. All rights reserved.

Citation

Sammer Siddiqui, Irfan Khan, Shamshad Zarina, Syed Ali. Use of the SYBR Green dye for measuring helicase activity. Enzyme and microbial technology. 2013 Mar 5;52(3):196-8