RNA-seq of WT2 Sorted_IgD-IgM-IgKint_splenocytes
Splenic CD19+CD3-IgD-IgK+ memory B cells from unimmunized mice were obtained by FACS. DNA was purified by ethanol precipitation. The JH4 intron region was amplified by PCR using the VHB1-8 framework three primer forward primer (5-CAGCCTGACATCTGAGGACTC-3) and reverse primer (5-CTCCACCAGACCTCTCTAGACA-3). 2% E-gel with SYBR safe (Invitrogen) were used to extract the JH4 amplified product, corresponding to a fragment of around 650bp in length and sequenced by Illumina MiSeq Nano using paired-end sequencing.