GSM3453680: 24hr_donor1; Homo sapiens; Hi-C
Naïve CD4 T cells were fixed by culture medium containing 1% formaldehyde for 5 minutes at room temperature and quenched by Tris (pH8.0). Nuclei were then isolated and subjected to SDS permeabilization followed by TritonX-100 to quench SDS. After that, nuclei pellets were subjected to HindIII digestion and re-ligation, at which step the biotin-dCTP was induced into the de novo ligation sites. Newly re-ligated genomic DNA were purified and sheared into 200bp fragments and fragments containing biotin-labeled de novo ligation sites were purified by M280 streptavidin beads (Invitrogen) and subjected to library preparation. DNA fragment was subjected to end-repair and biotin-remove by a master mix containing T4 DNA ligase buffer, dNTP mix, T4 PNK, T4 DNA Polymerase I and DNA polymerase I Large (Klenow) Fragment. Then dATP was attached to fragment end by Klenow exo minus enzyme. Resulted DNA fragment was ligated with Illumina indexed adaptors and amplified using Illumina PCR primer 1 and 2.
GEO
GSM3453680