title: GSM5469228: Spleen_3_WT2_JH1; Mus musculus; OTHER
library_construction_protocol: B cells were isolated from mice aged 5-7 weeks. Splenic B cells were isolated using B220 magnetic beads (Miltenyi Biotec) or by FACS sorting. B220+ IgM- cells were purified via FACS sorting from bone marrow. Genomic DNA samples were obtained using phenol-chloroform extraction of whole cell lysates. HTGTS-Rep-seq libraries were prepared as previously described (Hu et al., 2016; Lin et al., 2016). Briefly, genomic DNA was sonicated and subjected to LAM-PCR using biotinylated JH1 or JH3 bait primers (Lin et al., 2016). LAM-PCR products were purified using Dynabeads MyONE C1 streptavidin beads (Life Technologies, 65002) and ligated to bridge adaptors. Adaptor-ligated products were amplified by nested PCR with indexed JH1/JH3 primers (8-bp indexes were unique to each sample) and primer annealed to the adaptor. PCR products were further tagged with Illumina sequencing adaptor sequences and size-selected via gel extraction. Libraries were sequenced by paired-end 300-bp sequencing on an Illumina MiSeq by the Yale Center for Genome Analysis.