GSM1539056: iPSC_CTL1R-C3 (GDB-339-1_S) ChIP-seq input 1; Homo sapiens; ChIP-Seq
Genomic DNA was extracted using the DNeasy Blood and Tissue Kit (Qiagen) according to manufacturer specifications. About 10^8 iPSCs have been cross linked for 10 minutes in PBS containing 1% formaldehyde at room temperature. Crosslinking was quenched adding Glycine to a final concentration of 125mM for 5 minutes at room temperature. Cells nuclei were lysed using ChIP lysis buffer (1% SDS, 10mM EDTA, 50mM Tris-HCl, pH 8.1) and sonicated using the Covaris Sonicator (see sample description) to obtain a sheared chromatin DNA bulk of 250 bp. Sheared chromatin was incubated with GTF2I (15ug) or LSD1 (10ug) antibodies overnight rotating at 4 degree. The day after the antibody was recovered using Dynabeads Protein G (Life Technologies) and washed 4 times using washing buffers. Finally DNA was eluted using SDS containing elution buffer and decrossliked overnight at 65 degree. The third day decrosslinked DNA was digested using proteinase K for 60 minutes at 45 degree and purified using DNA-purification columns (Qiagen).
GEO Accession
GSM1539056