Bulk BCR repetoire requencing of homo sapiens:  adult with myasthenia gravis
RNA (250 ng) was reverse-transcribed into cDNA using a biotinylated oligo dT primer. An adaptor sequence was added to the 3' end of all cDNA, which contains a universal priming site and a 17-nucleotide unique molecular identifier (UMI). Products were purified using streptavidin-coated magnetic beads followed by a primary PCR reaction using a pool of primers targeting the IGHA, IGHD, IGHE, IGHG, and IGHM regions. The immunoglobulin-specific primers contained tail sequences for later Illumina adaptor addition. PCR products were then purified using AMPure XP beads. A secondary PCR was then performed to add the Illumina P5 adaptor to the constant region end and a sample-indexed P7 adaptor to the universal adaptor end. The number of secondary PCR cycles was tailored to each sample to avoid entering plateau phase, as judged by a prior quantitative PCR analysis. Final products were purified, quantified with a TapeStation (Agilent Genomics) and pooled in equimolar proportions, followed by high-throughput 2x300 base-pair paired-end sequencing with a 20% PhiX spike on the Illumina MiSeq platform according to manufacturers recommendations, except for performing 325 cycles for read 1 and 275 cycles for read 2.