GSM3259628: Il2ramut/mut_SPCD4+thym_input_thymus; Mus musculus; ChIP-Seq
Cross-linked DNA was lysed in Farnham lysis buffer (5mM PIPES pH 8.0, 85 mM KCl and 0.5% NP-40), fragmented in RIPA buffer (1% NP-40, 0.5% sodium deoxycholate, and 0.1% SDS in 1xPBS) using Bioruptor (Diagenode), and incubated overnight at 4_ with 100 µl of DynaBeads M-280 Sheep anti-Rabbit IgG magnet beads (Invitrogen) preincubated with 20 µg of anti-SATB1 antibody (Abcam, ab70004).  Beads were washed 5 times with LiCl wash buffer (100 mM Tris pH 7.5, 500 mM LiCl, 1% NP-40 and 1% sodium deoxycholate) and one time with 10mM Tris/0.1M EDTA. SATB1-bound DNA was then eluted from the beads and reverse cross-linked by incubating the beads pellet in 200 µl of IP elution buffer (1% SDS and 0.1 M NaHCO3) at 65_ overnight and further purified with MinElute PCR Purification Kit (QIAGEN) The library was prepared using Accel-NGS 2S Plus DNA library kit according to the manufacturer's instructions