GSM7663379: SEnd-seq of Nascent RNA without rpoc-his induction, replicate 2; Mycobacterium tuberculosis; OTHER
The cell culture was mixed with an equal volume of frozen 2× crush buffer (20 mM Tris-HCl pH 7.8, 10 mM EDTA, 100 mM NaCl, 1 M Urea, 25 mM NaN3, 2 mM β-mercaptoethanol, 10% ethanol, 0.4% NP40, and 1 mM PMSF) to stabalize RNAP complex. After immobilization with Ni-NTA agarose and thorough washing, the RNA was extracted through elution with 300 mM of imidazole. The RNA was further treated with DNase and cleaned with phenol-chloroform before library preparation. SEnd-seq strategy was used to prepare the sequencing libraries.