Electrophoretic mobility shift assays for with WhiB7

Published: 13-04-2021| Version 2 | DOI: 10.17632/fh9639t5sc.2
Elizabeth Campbell


EMSA on the whiB7 promoter with Mycobacterium tuberculosis RNAP holoenzyme, CarD and RbpA with and without WhiB7.


Steps to reproduce

Mtb holo RNAP with and without WhiB7 were assembled as described in the preparation of holoenzymes for cryo-EM, except the concentrations of Mtb holo RNAP were 1 µM, CarD 5 µM, RbpA 5 µM, and the DNA 100 nM. Reactions were run on a 4.5% polyacrylamide native gel (37.5:1 acrylamide:bis-acrylamide) in 1x TBE (89 mM Tris, 89 mM boric acid, 1 mM EDTA) at 4 ℃. The gel was stained with Gel-Red (Biotium).