GSM8727508: double negative DRGs; Mus musculus; RNA-Seq
DRG neurons are dissected and dissociated and loaded to FACS sorting. Sorted cells are loadded onto 10x Genomics Each sample was processed using the Chromium Next GEM Single Cell 3' Reagent Kits, v3, single index (10x Genomics). Chromium Single Cell 3' GEM, Library & Gel Bead Kit v3 (10x Genomics; PN-1000092), the Chromium Chip B Single Cell Kit (10x Genomics; PN- 1000073), and the Chromium i7 Multiplex Kit v2 (10x Genomics; PN-120262). Briefly, single cells were partitioned into Gel Beads-in-emulsion (GEMS) using the Chromium X instrument (10x Genomics). The presence of poly(dT) primer enabled the creation of full-length cDNA from poly-adenylated mRNA. cDNA was amplified to generate adequate mass for library construction. Sequencing libraries were prepared using end repair, A-tailing, adaptor ligation followed by PCR.