GSM4568535: TCF7L2_1; Homo sapiens; ChIP-Seq
Cells were washed 2x with ice cold PBS and then scraped in 1mL PBS into a 1.5mL Eppendorf tube. Fixed cells were spun (200xg, 10 minutes) and treated with lysis buffer according to the MAGnify Chromatin Immunoprecipitation kit protocol (150 μL/3 million cells assuming 15x106 cells/flask). Chromatin was sheared using a Bioruptor UCD2000 sonicator with the following settings: 12x cycles of 30 seconds sonication with 90 seconds cooling. Chromatin was sonicated in 1.5 mL eppendorf tubes, and the volume of cell lysate in each tube during sonication was kept at 150 μL. Sheared chromatin in lysis buffer was stored at -80°C prior to ChIP. ChIP reactions were carried out with MAGnify Chromatin Immunoprecipitation System (ThermoFisher) reagents as per manufacturer's instructions. Antibodies used in ChIP reactions were: 1 μg anti-TCF4 (Santa Cruz: sc-8631 X), 3 μL anti-H3K4me3 (Active Motif; 39159), 5 μg anti-H3K27ac (Active Motif; 39133). ChIP-seq Libraries were created with TruSeq ChIP Library Preparation Kit (Illumina) with DNA size selected and quality controlled by Pippin Prep size selection (Sage Science). Libraries were read with HiSeq SR Cluster v4 kit (Illumina) on a HiSeq 2500 sequencer with 50-bp single-end reads.
GEO Accession
GSM4568535