GSM7047529: Hi-TrAC_CTCF-AID_Tir1_bio2_tech3; Homo sapiens; OTHER
For Hi-TrAC, genomic DNA was purified by Phenol-Chloroform extraction. For RNA-seq, RNA was extracted with QIAzol lysis reagent (QIAGEN) and RNeasy mini kit (QIAGEN). For ChIP-seq, genomic DNA was purified with QIAGEN MinElute Reaction Cleanup Kit. For Hi-TrAC, repair gaps of purified genomic DNA with T4 DNA polymerase. Remove free bridging linker using AMPure XP beads. Then, digest purified DNA with MluCI and NlaIII restriction enzymes. Enrich biotin-labeled DNA fragments with streptavidin beads. Perform adapter ligation on beads. After washing the beads, amplify the library by PCR with illumina multiplexing indexed primers. For RNA-seq, libraries were constructed following Smart-seq2 protocol (Picelli et al., 2014). For ChIP-seq, DNA was purified, then end-repaired using End-It DNA End-Repair kit (Epicentre). A-tailing was performed with Klenow Fragment (3'->5' exo-) at the presence of dATP. Universal adapters were then ligated. The libraries were then amplified by PCR using indexed primers.