IgKL BCR repertoire of pRD patient
RNA was amplified using a commercially available multiplex primer mix covering the BCR heavy chain and/or kappa/lambda chains (iRepertoire Inc.). Briefly, reverse transcription and subsequent PCR amplification (RT-PCR1) is performed using Qiagen OneStep RT PCR mix (Qiagen). The cDNA is selected and unused primer is removed by SPRIselect bead selection (Beckman Coulter) followed by a second round of amplification performed with a pair of primers that are specific for communal sites engineered onto the 5 end of the C- and V- primers used during RT-PCR1. The final constructed library includes Illumina sequencing adapters and a 6-nucleotide internal barcode associated with the C-gene primer.