title: GSM3711512: KO3_CD3CD28; Mus musculus; RNA-Seq
library_construction_protocol: Sorted CD62LhiCD44lo WT and Morrbid-/- splenic CD8 T cells were stimulated for 6 hours at 1x10^6 cells/mL using 1μg/mL plate-bound αCD3 (145-2C11), 1μg/mL soluble αCD28 (37.51) with and without 20ng/mL recombinant IFN-1 (Biolegend). RNA was harvested using a combination of TRIzol (Invitrogen) and RNeasy Mini columns (QIAGEN). TRIzol-chloroform extractions were precipitated with ethanol, applied to RNeasy Mini columns, and processed according to the manufacturer's instructions. Sequencing libraries were generated using the SMARTer Stranded Total RNA Sample Prep Kit Hi Mammalian kit (Takara) using 100ng of total RNA. Library quality and concentration were assessed by TapeStation and qPCR respectively. Libraries were paired-end 75 base-pair sequenced on the NextSeq550 machine (Illumina).
attributes.tag: GEO Accession
attributes.value: GSM3711512