GSM3723570: cMyc+ GC-B CELL_FL110216_DC2_C12; Mus musculus; RNA-Seq
Spleens were crushed and filtered with a 70µM mesh strainer. Harvested splenic B-cells were enriched by CD43 negative selection using MACS LS Columns (Miltenyi Biotec). Single-cell suspensions were stained with antibodies and sorted by flow cytometry. The Fluidigm C1 automated microfluidic system was used to randomly capture 96 individual cells onto the 5–10 μm C1 IFCs (Fluidigm) for mRNA-seq. The SMARTer Ultra Low Input RNA kit for Sequencing (Takara Bio) was used to reverse transcribe poly(A) RNA and amplify cDNA. Amplified cDNA at a concentration of 0.10–0.3 ng/μL was used for preparing libraries with the Nextera XT DNA Library Preparation Kit (Illumina).