BCR-Seq of Homo sapiens: adult
PBMCs were isolated using the Lonza lymphocyte separation medium and cryopreserved until ready for processing. PBMCs were then thawed, incubated with DNaseI, washed, lysed in 1 ml TRIzol, and frozen at -20C. RNA was isolated using the Qiagen RNeasy kit and reverse-transcribed with SuperScript IV (Invitrogen). IgG + IgA variable heavy chain transcript was amplified in one reaction, and IgM variable heavy chain transcript was amplified in a second reaction.