GSM3665390: PAX6D INPUT1; Homo sapiens; ChIP-Seq
the cultures were digested with TrypLE to become single cells and then were crosslinked using 1% formaldehyde provided by the kit.  The cells were incubated at room temperature for 10 mins in a chemical fume hood. After incubation, the cells were treated with glycine solution for 5 mins at room temperature. After washing with cold PBS twice, the cells were lysed with the lysis buffer in the presence of protease inhibitors and MNase (provided by the kit to digest DNA). They were then sonicated (three 20-second pulses at 3 watts power) to yield DNA fragments of about 150bp to 1000bp. After taking 10 µl samples as an INPUT the rest samples were incubated with 1 µg FLAG antibody in 100 µl IP buffer provided by the kit overnight at 2-8 °C. The pull-downs were harvested by ChIP Grade Protein A/G Magnetic Beads. The DNAs were recovered from the INPUTs and IPs RNA libraries were prepared for sequencing using standard Illumina protocols