RNA-Seq of mus musculus: splenic B cells
Following cold PBS perfusion, mouse tumor tissue or spleen was minced and enzymatically dissociated at 37C for one hour. The CD45+, CD3- and CD19+ cells were FACS enriched using indicated antibodies and then centrifuged. After centrifugation, the supernatant was discarded. The cells were washed, resuspended in PBS, and counted. Only samples with cell viability exceeding 90% were processed further. Single-cell suspensions were loaded onto a microfluidic chip, and libraries were constructed following the manufacturer's protocol for subsequent sequencing on an Illumina HiSeq 4000 platform.