GSM2251959: Sig70 in E. coli, exponential aerobic growth, no rifampicin applied replicate 1 (run 423); Escherichia coli; ChIP-Seq
The samples were sonicated for a total of 16 minutes and 50 uL protein A/G sepharose. All reactions are performed in an Eppendorf ThermalMixerR while being mixed at 1.4k x rpm. After each reaction, the sepharose was washed with 1 mL of each of the following buffers, leaving 50-100 uL buffer above the sepharose in-between each wash : 1x TE : 10mM Tris-HCl, pH 8.0 + 1 mM EDTA ; 1 x High Salt - 1M NaCl (1% Triton X-100, 50 mM HEPES, pH 8.0, 2 mM EDTA, 0.1% sodium deoxycholate and 1M NaCl) ; 1x Wash Buffer 2 (1% Triton X-100, 50 mM HEPES, pH 8.0, 2 mM EDTA, 0.1% sodium deoxycholate and 0.5M NaCl) ;1x Wash Buffer 3 (1% Triton X-100, 50 mM HEPES, pH 8.0, 2 mM EDTA, 1.0% sodium deoxycholate, 0.25 M LiCl, 1% NP-40, and 10 mM Tris-HCl, pH 8.0) ; 1x TE : 10mM Tris-HCl, pH 8.0 + 1mM EDTA. The ChIP-exo experiments were generated by following the steps presented by Rhee and Pugh, 2011.