ChIP-Atlas: SRX1993427

Antigen

Cell type Class: Placenta
Cell type: Placenta
MeSH Description: A highly vascularized mammalian fetal-maternal organ and major site of transport of oxygen, nutrients, and fetal waste products. It includes a fetal portion (CHORIONIC VILLI) derived from TROPHOBLASTS and a maternal portion (DECIDUA) derived from the uterine ENDOMETRIUM. The placenta produces an array of steroid, protein and peptide hormones (PLACENTAL HORMONES).

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: Viable cells were counted using hemocytometer and crosslinked with 4% paraformaldehyde (Electron Microscopy Sciences, PA) for 10 minutes at room temperature before nuclear isolation. Cross-linked chromatin was prepared and sonicated using Bioruptor UCD-200 in ice water bath to generate DNA fragments 200-300 bp in size. Twenty micrograms of each antibody preparation, or 20 µg IgG for mock pulldown controls, were incubated with chromatin prepared from nuclei of approximately 5 million cells. DNA was released and quantitated using Qubit 2.0 (Life Technologies) with dsDNA HS Assay kit (Life Technologies, Q32854), and 15 ng of DNA was used to generate libraries for Illumina sequencing. ChIP-seq libraries were generated using KAPA LTP Library Preparation Kits (Kapa Biosystems, KK8232) to yield two independent ChIP replicates for the antibody. We also generated libraries from sonicated genomic input DNA from the same chromatin preparations as controls. Libraries were bar-coded with Bioo Scientific index adapters and sequenced to generate 15-23 million reads per duplicate sample using the Illumina Hi-Seq 2000 instrument at the University of Illinois W.M. Keck Center for Comparative and Functional Genomics according to manufacturer’s instructions.