ChIPseq control (input) for KN99deltaH1
Freeze-dried cell pellets were broken into fine powder with 0.5 mm zirconia/silica beads in a FastPrep 24 homogenizer (MP Biomedicals) without buffer for three cycles (50s at 8m/s followed by 90s chilling on ice) and then with 250 l of ChIP lysis buffer for another four cycles. The lysate was recovered on ice. Lysate was then diluted to 1 ml with ChIP lysis buffer without detergents and transferred to a milliTUBE 1 ml with AFA Fiber. Samples were sheared with the Covaris E220 Focused ultrasonicator for 13 minutes at 5C. The sheared chromatin was centrifuged at 21000 g for 15 min at 4C to remove insoluble cellular debris. Twenty microliters of sheared chromatin suspension were saved as Input DNA control and stored at 20 C.