BCR-seq of captive Artibeus jamaicensis: adult male spleen
RNA was extracted from splenic tissue with the Quick-RNA/DNA miniprep plus (Zymo #D7003) kit. cDNA was prepared using the SuperScript III First Strand Synthesis System (Invitrogen #18-080-051). BCR sequences were amplified from cDNA by PCR using a pool of forward primers targeting the beginning of IGHV genes and a reverse primer targeting the CH1 region of either IGHM or IGHG. PCR was performed using AmpliTaq Gold 360 DNA Polymerase (Applied Biosystems #43-988-23). Illumina sequencing libraries were prepared from cleaned PCR reactions using the NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB #E7103S) and NEBNext Multiplex Oligos for Illumina (NEB #E7600S). Indexed IGHM and IGHG libraries were pooled for each bat and sequenced on an Illumina MiSeq (2 x 300 bp kit).