DNBSEQ-G400 paired end sequencing; Raw reads: 6M_CORTEX_10X_R2_G2
Nuclei were sorted by FACS to obtain 700-1000nuclei/ul suspension. Nuclei was loaded to Chromium Next GEM Chip G (PN-1000120, 10x Genomics) aiming for a target recovery of 5000-7000 nuclei according to Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 protocol(CG000204 • Rev D, 10x Genomics).RNA extraction, cDNA construction and amplification was done according to Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 protocol(CG000204 • Rev D, 10x Genomics). Gel Beads-in-emulsions(GEMs) containing single nuclei were generated by loading Single Cell 3ʹ v3.1 Gel Beads, a Master Mix containing cells, and Partitioning Oil onto Chromium Next GEM Chip G (PN-1000120, 10x Genomics).The poly-adenylated mRNA was then captured and barcoded by primers including, TruSeq Read 1,10xBarcode, UMI,Poly(dT)VN) on Single Cell 3ʹ v3.1 Gel Bead (PN-1000122, 10x Genomics).
Nuclei were sorted by FACS to obtain 700-1000nuclei/ul suspension. Nuclei was loaded to Chromium Next GEM Chip G (PN-1000120, 10x Genomics) aiming for a target recovery of 5000-7000 nuclei according to Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 protocol(CG000204 • Rev D, 10x Genomics).RNA extraction, cDNA construction and amplification was done according to Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 protocol(CG000204 • Rev D, 10x Genomics). Gel Beads-in-emulsions(GEMs) containing single nuclei were generated by loading Single Cell 3ʹ v3.1 Gel Beads, a Master Mix containing cells, and Partitioning Oil onto Chromium Next GEM Chip G (PN-1000120, 10x Genomics).The poly-adenylated mRNA was then captured and barcoded by primers including, TruSeq Read 1,10xBarcode, UMI,Poly(dT)VN) on Single Cell 3ʹ v3.1 Gel Bead (PN-1000122, 10x Genomics).