| Assay Method Information | |
| | Helicase Assays |
| Description: | Standard helicase reaction mixtures (20μl) containing 5 fmol of the indicated TP-G4 or OX-1-G2' DNA substrates or forked duplex DNA substrate (0.25 nM DNA substrate concentration), 1mM ATP, and the indicated concentrations of the specified helicase were as previously described for FANCJ [Wu et al., Mol. Cell. Biol., 28:4116-4128] and DDX11 [Wu et al., J. Biol. Chem., 287:1007-1021]. Helicase reactions for XPD and DinG were performed in buffer that contained 20mM Tris-HCl (pH 8.0), 10mM KCl, 1mM dithiothreitol, 5mM MgCl2, and 100 μg/ml BSA. Reactions were performed at 37 °C for 15 min in the presence of 1 mM ATP, the DNA substrates mentioned above, and the indicated helicase protein concentrations. Forthe unimolecular Poly(A) Zic1-G4 DNA substrate, helicase reactions were performed in reaction buffer containing a 20-fold excess of peptide-nucleic acid complementary oligonucleotide [Giri et al., Nucleic Acids Res., 39:7161-7178]. G4 helicase reactions were terminated by th |
| Affinity data for this assay | |
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