Single cells derived from a C57BL mouse single Peyer's patch were sorted as either germinal center cells (B220+ CD138- FAS+ GL7+ CCR6- IgA+) or subepithelial dome cells (B220+ CD138- FAS+ GL7- CCR6+ IgA+). Amplified cDNA sequence of the B cell receptor was sequenced by Sanger sequencing and analyzed.. IgA B cell receptor sequencing

Single cells derived from a C57BL mouse single Peyer's patch were sorted as either germinal center cells (B220+ CD138- FAS+ GL7+ CCR6- IgA+) or subepithelial dome cells (B220+ CD138- FAS+ GL7- CCR6+ IgA+). Amplified cDNA sequence of the B cell receptor was sequenced by Sanger sequencing and analyzed.Single cell suspension of a single Peyer's patch was analyzed by FACS and single cells were sorted according to the markers described above into lysis buffer. cDNA was prepared using random primers. Specific nested PCR of a IgA antibody sequence was performed (with a constant primer and a mix of primers in the variable region). Sequencing was performed by Sanger sequencing. Sequences were validated using snap gene and IgBlast.