title: GSM3370177: WT_HPC7_Capture-C_Rep2; Mus musculus; OTHER
library_construction_protocol: E14.5 fetal liver and adult bone marrow were dissected and HSCs were purified using cell surface marker: Lin-Sca1+cKit+CD135-. Half million FACS-sorted cells were cross-linked, lysed and nuclei were digested using DpnII (R0543T, NEB) at 37°C with shaking at 1400 rpm overnight. The digested chromatin was proximally ligated with 100 U T4 DNA ligase (10799009001, Roche) at 16°C for 8 hrs with slow rotation. The samples were de-cross-linked followed by purification using phenol-chloroform extraction and precipitated using 70% ethanol. The purified 3C DNA was was sonicated to 200~300bp using Covaris S220 ultra-sonicator (6 min; duty cycle, 10%; intensity, 5; cycle per burst, 200). Sequencing libraries were constructed using NEBNext Ultra Kit (E7370, NEB). The libraries were size-selected using AMPure XP beads (A63881, Beckman Coulter). Oligonucleotide capture was performed using the SureSelect XT2 protocol (G9621A, Agilent). The libraries were quantified with Qubit, Agilent Bioanalyzer 2100 and KAPA qPCR (KK4844, Kapabiosystems). The captured libraries were sequenced with 150 bp paired-end reads on an Illumina HiSeq platform. Libraries constructed using NEBNext Ultra Kit (E7370, NEB). The libraries were size-selected using AMPure XP beads (A63881, Beckman Coulter). The libraries were quantified with Qubit, Agilent Bioanalyzer 2100 and KAPA qPCR (KK4844, Kapabiosystems). The captured libraries were sequenced with 150 bp paired-end reads on an Illumina HiSeq platform.
attributes.tag: GEO Accession
attributes.value: GSM3370177