GSM5777034: A3B_SPIseq_siNT_E2_rep1; Homo sapiens; OTHER
SPI protocol was adapted from the ChIP protocol used by Myer's Lab at HudsonAlpha (protocol version v011014) which contributed to the ENCODE project. Except that: i), cells were lysed in ChromaTrap lysis buffer that designed for sonication (Porvair Science, 100001), and chromatin fragmentation was carried out on a Covaris E220 instrument with optimised settings for ChIP purpose; 2), ChIP-grade protein A/G beads (Pierce 26162) were used to perform immuno-precipitation experiments; 3), Flag-M2 (Sigma F1804) antibody was used, and the following buffer was used for chromatin binding and washing: 100 mM Tris pH 7.5; 500 mM LiCl; 1% NP-40; 1% sodium deoxycholate; 4), Following ChIP, the immunoprecipitated complex were subjected to denaturing and adapter ligation following the protocol reported by the developers of SPI-seq (PMID: 23249883). Library was constructed following the protocol reported by the developers of SPI-seq (PMID: 23249883).