GSM9384277: IgG_3 (RNA-seq); Mus musculus; RNA-Seq
Spleen and dLN from influenza infected mice were homogenized, red blood cells lysed with ACK Lysis buffer (150mM NH4Cl, 10mM KHCO3 and 0.5M EDTA (pH 7.3)). Lymphocytes from influenza infected mice were stained for cell sorting. For each sample, 1,000 cells were sorted into RLT buffer (Qiagen 79216) containing 1% 2-Mercaptoethanol. RNA was isolated using Zymo Quick-RNA MicroPrep Kit (Zymo Research, R1051). Synthesis of cDNA was performed using SMART-Seq v4 Ultra Low Input RNA Kit (634894; Takara Bio) kit. Final libraries were generated using 200 pg of cDNA as input for the NexteraXT kit (Illumina, FC-131-1024) with 12 cycles of PCR amplification.