IGHV-Seq of mus musculus fetal liver FCRL6- pro B cells
FCRL6+ and FCRL6- pro B (AA4.1+Lin-CD43+B220+CD19-IgM-) cells, derived from the fetal livers of embryonic day 18 and bone marrow of adult 8-12 week old mice, were sorted by flow cytometry from BALB/cJ strain mice. Single cell suspensions from leg bones were pooled from 5-10 adult male and female mice and 7-10 fetal livers were pooled from embryonic day 18 fetuses isolated from pregnant mothers according to the timing of vaginal plug formation. Cells were lysed in trizol, RNA was isolated and used to generate cDNA. Rearranged IGHV genes in these samples were amplified, barcoded, and sequenced.