GSM2330286: D3_INPUT; Pseudomonas aeruginosa; ChIP-Seq
Chromosomal DNA was sheared by sonication to an average size of 0.5-1 kb. After the removal of cell debris by centrifugation, 50 μl of each sample was removed to serve as an input control. The remaining samples were added to 50 μl of ANTI-FLAG M2 affinity resin (Sigma A2220) previously washed twice with TBS and twice with lysis buffer. After incubation at 4°C overnight, beads were pelleted and washed twice with TBS Tween 0.5% and three times with TBS. Elution was performed using the 3X FLAG peptide (Sigma F4799), as recommended. The recovered supernatants were placed at 65°C overnight to reverse the cross-links. The input samples were also incubated at 65°C overnight after the addition of 200 μl of TBS. Libraries preparation and sequencing were performed by the IMAGIF facility (I2BC, Gif sur Yvette), according to Illumina recommendations.
GEO Accession
GSM2330286