Electrophoretic mobility shift assays for with WhiB7

Published: 13 April 2021| Version 2 | DOI: 10.17632/fh9639t5sc.2
Contributor:
Elizabeth Campbell

Description

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

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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).

Categories

RNA Transcription

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