GSM1436629
GSM1436629: WCE; Mus musculus; ChIP-Seq
The mouse hematopoietic stem and progenitor cell population for ChIP was isolated from E14.5 foetal livers from C57BL/6 mice by fluorescence-activated cell sorting based on the following cell surface marker profile: lineage (Ter119, B220, CD5, CD3, Gr1) negative, c-Kit+ and Sca1+. Similarly, adult bone marrow hematopoietic stem and progenitor cells for RNA-seq were sorted based on lineage (Ter119, B220, CD19, Mac1, Gr1, CD2, CD3, CD8) negative, c-Kit+ and Sca1+ cell surface marker expression. Approximately 250,000 cells were used for each ChIP, whereas 30,000 to 100,000 cells were used to prepare RNA for the RNA-seq. For chromatin immunoprecipitation, formaldehyde cross-linked cells were sonicated in a Covaris sonicator.  A small fraction of sonicated material was retained as the WCE sample and the remainder was incubated with either an antibody against H3 (AbCam) or H3K27me3 (Millipore) overnight at 4°C. Immune complexes were purified by incubation with protein G beads (Life Technologies) at 4°C for an hour.  Cross-links were reversed by incubation at 65°C for 4 hours before DNA fragments were purified with the ChIP Clean and Concentrator kit (Zymo). Sequencing libraries were prepared using the TruSeq DNA Sample Prep Kit (Illumina) and sequencing was performed on a HiSeq2000 (Illumina).
GEO
GSM1436629
GEO Accession
GSM1436629