ChIP-Atlas: SRX7099289

Antigen

Cell type Class: Blood
Cell type: Hematopoietic Stem Cells
MeSH Description: Progenitor cells from which all blood cells derived. They are found primarily in the bone marrow and also in small numbers in the peripheral blood.

library_strategy: ATAC-seq
library_source: GENOMIC
library_selection: other
library_construction_protocol: For RNA-seq, HSCs (Lin-, cKit+, Sca1+, CD150+, CD48-) and Rps (Lin-, cKit+, Sca1+, C-150- CD48+) were sorted from a pooled sample of 2 male and 2 female 8-week old mice HSCs were purified from three or four biological replicates (composed of pooled WBM from three male and three female mice) of either Vav-CRE: Dnmt3a+/+; Tet2+/+ or Vav-CRE:Dnmt3afl/fl or + or Vav-CRE:Tet2fl/fl mice. Total RNA was isolated from the samples using the NucleoSpin RNA XS kit For RNA-seq, the SMARTer Ultra Low RNA kit (Clontech) was used to prepare the libraries from 3-5ng of total RNA. For ATAC-seq, HSCs (Lin-, cKit+, Sca1+, CD150+, CD48-),MPPs ( Lin-, cKit+, Sca1+, C-150-, CD48-) and Rps (Lin-, cKit+, Sca1+, C-1-0+, CD+8-) were sorted from a pooled sample of 2 male and 2 female 8-week old mice HSCs were purified from three or four biological replicates (composed of pooled WBM from three male and three female mice) of either Vav-CRE: Dnmt3a+/+; Tet2+/+ or Vav-CRE:Dnmt3afl/fl or + or Vav-CRE:Tet2fl/fl mice. For ATAC-Seq, NEBNext High-Fidelity 2X PCR Master MIX (NEB M0541L) along with Nextera custom primers were used for library amplification of the ATAC DNA(5-7 cycles of PCR as determined by qPCR). Libraries were again purified with Ampure XP beads and run on an Illumina Hiseq 3000 (PE2X150) or NovaSeq(PE2x150).