GSM6481107
Bead-bound cells were suspended by dig wash buffer with 1:50 dilution of anti-MLL1, RNAPII, YTHDC2 or H3K4me3 antibody incubated on a roating platform overnight at 4 °C.After removing primary antibodies, the corresponding secondary antibody was diluted 1:100 in dig wash buffer and cells were incubated for 60 min. Then cells were washed using dig wash buffer. A 1:100 dilution of pA-Tn5 adapter complex was prepared in dig-med buffer and incubated with cells for 1 h. Cells were washed with Dig-med buffer. Then cells were resuspended in tagmentation buffer (10mM MgCl2 in Dig-med Buffer) and incubated at 37 °C for 1 h. DNA was purified using phenol-chloroform-isoamyl alcohol extraction and ethanol precipitation. Sequencing was performed in the Illumina Novaseq 6000 using 150bp paired-end.