title: GSM7913094: Splenic CD19+ Ig transcript library, WT C57BL/6J sample 1; Mus musculus; OTHER
library_construction_protocol: CD19+ splenocytes were isolated 10.5-week-old 3 WT and 3 Mtv8KO C57BL/6J female mice. Red blood cell lysed splenocytes were labeled with microbeads conjugated to monoclonal anti-mouse CD19 antibodies (isotype: rat IgG2a) (Miltenyi Biotec, Bergisch Gladbach, Germany) and positively sorted as detailed by the manufacturer. RNA was isolated from isolated CD19+ splenocytes using guanidine thiocyanate extraction and CsCl gradient centrifugation. Immunoglobulin libraries were generated from 1 mg of CD19+ splenocyte RNA using the NEBNext Immune Sequencing Kit (New England Biolabs, Ipswich, Massachusetts, USA) according to manufacturer's instructions, specifically enriching for B cell receptor chains during the first PCR step. Six libraries were pooled in equimolar amounts and were sequenced by paired-end 300 bp sequencing on an Illumina MiSeq by The University of Chicago Genomics Facility.