GSM7697898: Trigeminal ganglion (TG) cells, replicate 2, scRNAseq; Mus musculus; RNA-Seq
Freshly collected trigeminal ganglia from wild-type untreated male C57black/B6 mice were treated by Librase (200ug/ml) and dispase II (300ug/ml) in Hanks for 60 min at 37 degree. Single cell susupension was generated by prituration of digested trigeminal ganglia. Suspension was filtered through 70um filter. Live cells were FACS sorted. Trigeminal ganglion cells were resuspended in DPBS supplemented with 0.04% w/v BSA. Library was performed according to the manufacter's instructions (single cell 3' v2 protocol, 10x Genomics). Briefly, GCs 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 poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an Ilumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the apropiated sized fragments were selected with SPRIselect reagent for subsequent library construction. During the library construction Ilumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.