GSM7596454: Keratinocytes, siLINC00941, input, rep2; Homo sapiens; ChIP-Seq
Cells were fixed using 1% formaldehyde at RT for 10 min, the reaction was quenched with 125 mM glycine at RT for 5 min. Cells were pelleted and then lysed in Swelling buffer (100 mM Tris, pH 7.5, 10 mM potassium acetate, 15 mM magnesium acetate, 1% Igepal, 1x cOmplete Protease Inhibitor Cocktail (Roche) and 1 mM AEBSF). Chromatin fragmentation was subsequently performed in RIPA buffer for 30 min using the S220 Focused-ultrasonicator (Covaris) in the Freq sweeping mode, intensity 8, 20% duty cycle and 200 cycles per burst. 100 µg fragmented chromatin underwent IP with 5 µg α