Extended Data Figure 1. Overview of BCR repertoire strategy.

a) Schematic diagram of the B cell receptor repertoire analysis strategy. b) Schematic diagram of the B cell receptor sequencing strategy. In the reverse transcription (RT) step, the primer anneals to the constant region of the B cell receptor mRNA to generate cDNA with a random 12 nucleotide barcode. This barcode can be computationally used to reduce PCR amplification biases after sequencing. The product is cleaned and PCR amplified using multiple primers binding to the FR1 region of the IgH genes along with a universal sequence complementary to the end of the RT primer. c) Gating strategy to flow sort B cell subsets from healthy donor PBMCs.