title: GSM8452796: PC_KO_4; Mus musculus; RNA-Seq
library_construction_protocol: Spleen from (un) infected mice were homogenized, red blood cells lysed with ACK Lysis buffer (150mM NH4Cl, 10mM KHCO3 and 0.5M EDTA (pH 7.3)). Splenocytes from LPS infected mice at 72hrs were stained for cell sorting. 1,000 Activated B cells and Plasma cells were sorted into RLT (Qiagen) + 1%BME, snap frozen and stored at -80°C till processing. Naive B cells were enriched from uninfected mice using CD43 (Ly-48) MicroBeads kit (Miltenyi Biotec) according to manufacturers protocol and 1,000 cells resuspended in RLT (Qiagen) + 1%BME, snap frozen and stored at -80°C till processing. Total RNA was purified from 1,000 enriched Naïve B cells or sorted Activated B cells or Plasma cells using the Quick-RNA MicroPrep Kit (Zymo Research). cDNA was synthesized using the SMART-seq v4 cDNA synthesis kit (Takara). 200pg cDNA, the NexteraXT kit with the NexteraXT indexing primers (Illumina) was used to generate final libraries.