JW-KO1-Cmu_S6_L001_R1
To analyze the repertoire usage and SHM in V186.2 region DNA, RNA was extracted from splenic B cells for cDNA synthesis. Rearranged V186.2DJH-Cm and V186.2DJH-Cg1 cDNA was amplified using Phusion high-fidelity DNA polymerase (New England BioLabs) and a V186.2 leader-specific forward primer and a reverse primer specific for the Cm or Cg1 exon, followed by the second round of PCR using the same forward primer and a nest reverse primer tagged with an Illumina clustering adapter.  The amplified library was tagged with barcodes for sample multiplexing and analyzed by 300-bp pair ended sequencing (the Illumina Mi-Seq system).