Repertoire analysis on BCR genes

BALB/c mice were intravenously injected with 2 or 4 combination doses of empty PEG-Lip and subsequent OVA-PEG-Lip every 2 weeks. As a comparison group, BALB/c mice were subcutaneously injected with an initial 2 or 4 doses of an OVA/CFA emulsion and sequentially with an OVA/IFA emulsion every 2 weeks. At 3 days post last injection, spleens was harvested from the mice. Total RNA was isolated from the spleen cells using ISOGEN. Sequencing of the BCR genes in splenic B cells was performed using the unbiased gene amplification method with Adaptor-Ligation PCR by Repertoire Genesis Inc.