GSM7027219: mouse OE, 19d_rep2, scRNAseq; Mus musculus; RNA-Seq
Mice were deeply anesthetized and perfused transcardially with 20 ml of ice-cold phosphate buffered saline (PBS). Mice were then rapidly decapitated, and turbinate tissues were washed and immersed in ice-cold PBS. Tissues were minced into small pieces and disaggregated in 0.25% Trypsin-EDTA (1x) (GIBCO) with gentle agitation at 37 °C for 20 min. After adequate disaggregation, adding equal volume DMEM containing 10% Fetal Bovine Serum (FBS, GIBCO) to stop the disaggregation reaction then the disaggregated cells were washed with ice-cold PBS and filtered through a 40 μm strainer. 7-AAD (Invitrogen) was added to exclude dead cells. For fluorescent reporter-expressing mouse lines (PWK/PhJ*129P2/OlaHsd (OMP-ChR2-YFP), fluorescence-positive cells (mOSNs) were sorted. Cells were sorted into DMEM containing 10% FBS and centrifuged for 5 minutes at 300 g and resuspended with PBS. Single-cell sequencing libraries were prepared using the Chromium Single Cell 3′ Reagent Kit V3 (10x Genomics) according to the manufacturer's guidelines. Sequencing library fragments were examined using the Agilent High Sensitivity DNA kit (Agilent) and quantified via qPCR by the KAPA library quantification kit (Roche).