IGH repertoire sequencing from B cell popoulations in individuals with CBL LOF variants

IGH repertoires were sequenced from sorted B cell subset (transitional, naive and memory) in individuals with loss of function variants in the CBL gene. IGH were amplified from cDNA using isotype specific primers; IgM (all cell populations), IgG (memory) and IgA (memory). PCR amplificated was independent for each isotype and used a isotype specific primer (IgM, IgG or IgA) with a pool of IGHV forward primers. Samples were barcoded with Nextera indexes for pooling and sequenced on the Illumina MiSeq platform with 2x300bp paired end reads.