Illumina HiSeq 4000 paired end sequencing; RNA-seq of mouse myeloma cell line SP 2/0 cells compared to naive B cells
RNA-seq of mouse myeloma cell line SP 2/0 cells compared to naive B cells
Splenic lymphocytes from male 9-week old C57BL/6 mice were separated by Ficoll solution. B cells were sorted by B220 microbeads. SP 2/0 cells   (ATCC® CRL-1581™, https://www.atcc.org/products/all/CRL-1581.aspx) were collected after after cultured for 2 days in fresh medium. Cell samples were washed twice with PBS and 1 mL of TRIzol per 1 x 107 cell pellets was added and frozen in -80℃. Data were aligned using STAR with options --outFilterScoreMinOverLread 0.4 --outFilterMatchNminOverLread 0.4, against the C3H/HeJ genome as a reference (ftp://ftp.ensembl.org/pub/release-89/fasta/mus_musculus_c3hhej/dna/Mus_musculus_c3hhej.C3H_HeJ_v1.dna_sm.toplevel.fa.gz).  Total RNA from cells was isolated and purified using the RNeasy Mini Kit (Qiagen, Venlo, Netherlands). All RNA content in the samples was measured with a NanoDrop®ND-1000 spectrophotometer, and the RNA quality was assessed using RNA 6000 NanoChips with the Agilent 2100 Bioanalyzer (Agilent, Palo Alto, CA, USA). Libraries were prepared using the TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Gold (Illumina).