GSM7816210: Unstimulated FO B cells (Unstimulated), rep1; Mus musculus; RNA-Seq
RNA was extracted from cells using miRNeasy Micro Kit (Qiagen). BRB-seq(Alpern D et al.) was performed for library preparation with some following modifications. Barcoded dT primer(5'-GCCGGTAATACGACTCACTATAGGGAGTTCTACAGTCCGACGATCNNNNNNNNNNCCCCCCCCCTTTTTTTTTTTTTTTTTTTTTTTTV -3' ; (10)N=UMI, (9)C=cell barcode) was used for reverse transcription. Second Strand Synthesis Module (NEB, #E6111) was used for double-stranded cDNA synthesis. In-house MEDS-B Tn5 transposase (Picelli S et al., Sato et al.) was used for tagmentation and libraries were amplified by 10 cycles of PCR using Phusion High-Fidelity DNA Polymerase(Thermo Scientific, #M0530) with following primers(5' -AATGATACGGCGACCACCGAGATCTACACindexGTTCAGAGTTCTACAGTCCGA-3',5'-CAAGCAGAAGACGGCATACGAGATindex GTCTCGTGGGCTCGGAGATGT-3').