GSM5767772: DMR363_t0_induced; Bacillus subtilis; ChIP-Seq
To purify the DNA, 500 μl phenol:chloroform:isoamyl alcohol mix (Sigma) was added to the IP and WCE tubes, mixed vigorously and centrifuged for 10 min at 13,000 rpm (at room temperature). 450 μl of aqueous phase was collected and precipitated with 1 ml 100 % ethanol in the presence of 45 μl NaOAc (Sigma) and 1.2 μl of Glycoblue (Invitrogen) for 20 min at −20°C. DNA pellets were collected by centrifugation and resuspended in 100 μl EB (Qiagen) by vigorous shaking for 10 min at 55°C. Final purification was performed using a PCR purification kit (Qiagen), eluting the DNA in 50 μl EB. Prior to deep sequencing, the success of the IP was verified with qPCR. For deep sequencing, the DNA libraries were prepared by the Genomic Facility at CIG, UNIL, Lausanne. Briefly, the DNA was fragmented by sonication (Covaris S2) until the DNA was sheared to 220-250 bp. The Ovation Ultralow Library Systems V2 kit (NuGEN) including 15 cycles of PCR amplification was used to prepare the DNA libraries. 12-15 million sequence reads per sample were obtained on a HiSeq4000 (Illumina) with 150 bp read length.