HEK293T_NT_input_ChIPseq
Homo sapiens
Cells from one 10 cm dish (~1 x 107) of HEK293T cells treated with non-targeting siRNA (D-001206-14, Dharmacon) were used for each immunoprecipitation. The cells were cross-linked with 2 mM DSG Crosslinker in PBS for 30 min and then 1% formaldehyde in PBS for 20 min at room temperature. Then the cells were resuspended and lysed in lysis buffer (0.5% SDS, 10mM EDTA, 150 mM NaCl, 50 mM Tris-HCl pH 8.0), and they were sonicated with a Bioruptor Sonicator (Diagenode, Denville, NJ) for 30 times for 30 seconds at the maximum power setting to generate DNA fragments of ~150-500 bps. Crosslinking was reversed by overnight incubation at 65C. Input DNA were treated with RNase A and Proteinase K by incubation at 45C. DNA was purified using the QIAquick PCR purification kit (28106, Qiagen).
sample_name
NT_Input
cell_line
HEK293T