GSM5171689: MYC-IP from KO-cells - Rep1; Homo sapiens; ChIP-Seq
Cells were treated with 1% formaldehyde for 9 min at 37 °C. After cell lysis, nuclei were re-suspended in RIPA buffer (10 mM Tris/HCl pH 7.5, 150 mM NaCl, 1% NP40, 1% deoxycholic acid (DOC), 0.1% SDS, 1 mM EDTA), 6% of murine spike-in were added, and DNA was fragmented to a size <300 bp using a Branson sonifier. Chromatin was immunoprecipitated with 10-15 µg of corresponding antibody for 6 hours. Chromatin was eluted with 1% SDS and crosslinking was reverted overnight. For purification, chloroform/phenol extraction was used. NEBNext® Ultra II DNA Library Prep with Sample Purification Beads®. Purified DNA was end-repaired, A-tailed, ligated to Illumina adaptors, size-selected (200 bp) and purified with Sample Purification Beads. DNA fragments were amplified by 18 cycles of PCR and library size was tested with the Fragment Analyzer (Advanced Analytical).
GEO Accession
GSM5171689