title: GSM5983522: ES AS AC-TRIM33 rep1; Mus musculus; ChIP-Seq
library_construction_protocol: For GFP-TRIM33-chip experiment, GFP-TRIM33 mESCs were cross-linked with 1% formaldehyde at 37 °C for 10 min and quenched with 0.125 M glycine for 5 min at room temperature. Then cells were scraped and GFP-trap-beads assisted ChIP was performed and DNA were extracted using a ChIP assay kit (Millipore) as described. For SMAD2/3, PML or H3K27ac chip, 20 million mESCs were collected for cut&tag experiment followed manuals (Yeason) using anti-SMAD2/3, PML or H3K27ac antibody  to got enriched DNA. For library construction and sequencing, ChIP DNA samples were quantified and quality assessed by Qubit 2.0 and Agilent 2100. DNA fragments range from 200-600bp were selected constructed for ChIP-Seq library with TruSeq ChIP Sample Prep Kit (Illumina) according to manufacturer's instructions. After scraped from plate, embryonic stem cells were washed with PBS and incubated in lysis buffer.