ChIP-seq analysis of RpoS in Vibrio cholerae
ChIP-seq analysis of RpoS in Vibrio cholerae
The Vibrio cholerae O1 El Tor strain A1552 was isolated in 1992 from a traveller from Peru. Cells were grown in liquid LB medium, at 37 degrees with shaking, to an OD600 of 2.0. Immunoprecipitations were done as described by Haycocks et al (PLoS Genet. 2019. 15, e1008362) using derivatives of Vibrio cholerae strain A1552 using anti-FLAG antibodies (Sigma).  Libraries were prepared using immunoprecipited protein-DNA complexes immobilised with Protein A-sepharose. DNA fragments were then blunt ended, A-tailed and barcoded. Following elution of complexes from the Protein-A sepharose, crosslinks were reversed and barcoded libraries were amplified by PCR. The number of PCR cycles was determined empirically for each library. After amplification, library concentration was quantified using Qubit analysis and real time PCR.