title: Amplicon-Seq of mus musculus: B cells from adult female spleen and lymph nodes
design: Sorted B cells were lysed for RNA extraction. First-strand cDNA synthesis was performed on total RNA using IgM and IgG specific primers. Second-strand synthesis reactions were performed with primers tagged with unique molecular identifiers. Flow cell-adaptor sequences and offset were introduced into dsDNA products. Single indices for demultiplexing were added using the NEBNext Multiplex Oligos for Illumina. All PCR products were purified by ExoSAP-IT and SPRI beads. Bead-purified libraries were quality controlled on Agilent 2100 Bioanalyzer or 4200 TapeStation, and quantified by qPCR.