title: GSM8974722: Olfactory Epithelium, WT; Mus musculus; RNA-Seq
library_construction_protocol: Olfactory epithelia were dissected out and minced with spring scissors in HBSS on ice. The minced tissues were incubated with  10 U/ml disease, 230 U/ml collagenase type II and 77 μg/ml DNase I in HBSS for 30 min at 37°C. Filter the cell suspension through a 40 μm strainer and a 10 μm strainer. Washed with Hibernate-A, 10% FBS two times and finally, cells were resuspended with 0.04% BSA in PBS. Library construction was performed according to the manufacturer's instructions (single cell 3' reagent kit v3.1). Briefly, dissociated cells were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in emulsion (GEM). The ploy-A RNA from the cell lysate continued in every single GEM was retrotranscripted to cDNA, which contains an Illumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcode cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the appropriate sized fragments were selected with SPRIselect reagent for subsequent library construction. During the library construction, Illumina Read2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.