Use of a restriction enzyme-digested PCR product as substrate for helicase assays.
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.
Shin
Jae-Ho
JH
University of Maryland Biotechnology Institute, Center for Advanced Research in Biotechnology 9600 Gudelsky Drive, Rockville, MD 20850, USA.
Reeve
John N
JN
Kelman
Zvi
Z
D001105:Archaea / Q000201:enzymology; D001419:Bacteria / Q000201:enzymology; D004265:DNA Helicases / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D017013:Methanobacteriaceae / Q000201:enzymology; D016133:Polymerase Chain Reaction / Q000379:methods*; D014780:Viruses / Q000201:enzymology
D004262:DNA Restriction Enzymes; D004265:DNA Helicases