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Analysis of DNA binding activity of the intact and mutant BLMs using electrophoretic mobility shift assay. One nanomole of 5′-labelled ssDNA or dsDNA was incubated at room temperature for 20 min with 2.5 to 160 nM protein in unwinding buffer. Bound and free DNA were separated by electrophoresis through a non-denaturing 15% polyacrylamide and revealed by autoradiography.
