3C-HTGTS for Linear RAG Scanning Mediates Editing of Igκ Variable Region Repertoires
3C-HTGTS for Linear RAG Scanning Mediates Editing of Igκ Variable Region Repertoires
Cells were fixed with 2% formaldehyde, quenched with glycine, lysed in a lysis buffer containing: 150mM NaCl, 0.5% NP-40, 1% Triton X-100, 50mM Tris-HCl pH7.5, 5mM EDTA pH8.0. Nuclei were digested with NlaIII, ligated with T4 DNA Ligase, treated with Proteinase K and RNase A prior to DNA precipitation. v-Abl cell lines were cultured in RPMI1640 media with 15% FBS G1-arrest was performed by treating v-Abl cell lines with 3uM STI for 4 days Cells were fixed with 2% formaldehyde, quenched with glycine, lysed in a lysis buffer containing: 150mM NaCl, 0.5% NP-40, 1% Triton X-100, 50mM Tris-HCl pH7.5, 5mM EDTA pH8.0. Nuclei were digested with NlaIII, ligated with T4 DNA Ligase, treated with Proteinase K and RNase A prior to DNA precipitation. 3C-HTGTS libraries were prepared using linear-amplification PCR-mediated high-throughput genome-wide translocation sequencing protocol with indicated bait primers