GSM6123243: Hs_hESC_D21_CDX2_ChIP_seq_input1; Homo sapiens; ChIP-Seq
Experimentally replicated day 21 human pluripotent stem cell-derived islets were collected in stage 6 differentiation media (MCDB131 with 1.5 g/L NaHCO3, 1X GlutaMAX, 20 mM D-glucose, 2% BSA, 100 nM LDN-193189, 1 µM T3, 100 nm γ-Secretase Inhibitor XX, 10 µM ZnSO4, 10 µg/mL heparin, 1:200 ITS-X) containing 1.11% formaldehyde and fixed on a rocker for 15 minutes. The reaction was quenched for 5 minutes in 0.125 M glycine. Fixed human pluripotent stem cell-derived islets were washed in DPBS containing 0.5% NP-40, then once again in DPBS supplemented with 0.5% NP-40 and 1 mM PMSF, and subjected to ChIP using the ChIP-IT High Sensitivity Kit (Active Motif) with 30 μg of sheared chromatin and 6 μg anti-CDX2 antibody (A300-691A, Bethyl Laboratories). Libraries were constructed from purified DNA using the KAPA DNA Library Preparation Kit for Illumina (Kapa Biosystems). Input libraries were prepared from each experimental replicate using 10 ng of DNA purified immediately following shearing. Libraries were sequenced using NovaSeq 6000 (Illumina) and reads were trimmed afterwards to fit into corresponding analysis pipeline.