title: Single-cell transcriptome and antibody repertoire sequencing of bone marrow B cells from mice following influenza (H1N1 and H3N2) intranasal infections
library_construction_protocol: Female 6-8 weeks old C57BL/6 mice (Janvier) were infected with 200 PFU of PR8 (A/PuertoRico/8/1934 (H1N1)) or X31 (a reassortant virus that contains the six internal segments from PR8 and the glycoprotein-encoding segments (HA/NA) from strain A/Aichi/1968 (H3N2)) (LD50 = 1250 PFU) intranasally (i.n.). the tibia and femur bones were flushed in ice-cold RPMI containing 10% FCS buffer with 10 ng/mL IL6 (216-16, Peprotech). An RBC lysis step was performed in 4 mL ammonium-chloride-potassium lysis buffer for 1 min at room temperature and subsequently inactivated with 26 mL RPMI containing 10% FCS. Cells were isolated via facs following the aforementioned experimental conditions 10x genomics 5' protocol was performed as according to manufacturers 10x genomics 5' protocol was performed as according to manufacturers