GSM1930760: MCF-10A wildtype BRG1 ChIPseq Replicate1 Pulldown; Homo sapiens; ChIP-Seq
~1x107 parental MCF-10A cells were crosslinked with formaldehyde at room temperature for 10 minutes.  Then, the cells were lysed using lysis buffer A (50mM HEPES, 140mM NaCl, 1mM EDTA pH=8, 10% Glycerol, 0.5% NP-40, 0.25% Triton X-100), and the residual cytoplasmic protein was removed using lysis buffer B (10mM Tris-HCl pH=8, 200mM NaCl, 1mM EDTA, 1mM EGTA).  The nuclear fraction was released using lysis buffer C (10mM Tris-Hcl pH=8, 100mM NaCl, 1mM EDTA, 1mM EGTA, 0.1% Sodium Deoxycholate, 0.5% N-lauroylsarcosine).  The chromatin was then sheared by using a Bioruptor instrument in high setting, 30' on , 30' off for 5 minutes for 5 cycles. The pull-down was performed by using the BRG1 antibody (Santa Cruz #G-7).  Samples were washed three times with RIPA buffer (Tris-HCl pH=8, 150mM NaCl, 1mM EDTA, 1% NP-40, 0.25% Sodium deoxycholate, 0.1% SDS) and were eluted. The pull-down and input control sequencing libraries were generated by using the NEXTflex Rapid DNA Sequencing Kit (Bioo Scientific #5144-02) and were sequenced by using SE100 reads with a HiSeq 2000 instrument.
GEO Accession
GSM1930760