Published online 13 January 2005
Methods Online |
Use of a restriction enzyme-digested PCR product as substrate for helicase assays
University of Maryland Biotechnology Institute, Center for Advanced Research in Biotechnology 9600 Gudelsky Drive, Rockville, MD 20850, USA 1 Department of Microbiology, The Ohio State University Columbus, OH 43210, USA
*To whom correspondence should be addressed. Tel: +1 301 738 6294; Fax: +1 301 738 6255; Email: kelman{at}umbi.umd.edu
Received November 1, 2004. Revised December 1, 2004. Accepted December 14, 2004.
DNA helicases play essential roles in many cellular processes. The currently available techniques to generate substrates for helicase assays are fairly complicated and need some expertise not available in all laboratories. Here, a PCR-based method to generate a substrate for a helicase assay is described, and its application for several archaeal, bacterial and viral enzymes is demonstrated.
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