9086-J70-run2
RNA was extracted from frozen PBMCs in TRIzol LS according to the manufacturers instructions. cDNA was synthesized from extracted RNA using SuperScript IV (SSIV) Reverse Transcriptase. Variable regions of heavy chains (of the IgM, IgG, and IgA isotypes) were amplified separately for each animal and time point. After purification, DNA concentrations were quantified using Quant-iTTM dsDNA Broad-Range Assay Kit. Two sequencing runs on two separate PCR amplification products were performed on a MiSeq sequencer (paired-end sequencing, 500 cycles, 2250).