title: GSM7114702: day14, scRNAseq; Homo sapiens; RNA-Seq
library_construction_protocol: Nuclei from cultured neurons were isolated as detailed in 10x Genomics demonstrated protocol CG000375 Rev A with modifications. Cells were washed with cold 1X DPBS then 500 uL of NP-40 Lysis Buffer was added and cells were gently scraped off the dish while on ice, transferred to a 1.5 mL tube, and incubated 5 min on ice. Cells were centrifuged at 500 x g for 5 min at 4C and 500 uL 1X PBS + 1% BSA + DAPI + 0.5U/uL Protector RNAse Inhibitor (Roche) was added. Cells were spun again, gently resuspended 5X in 0.1X Lysis Buffer, and incubated 2 min on ice. After incubation, 1 mL of wash buffer was added and cells were immediately spun at 500 x g for 5 min at 4C. Nuclei were resuspended in <100 uL of Diluted Nuclei Buffer and counted using Countess FL II aiming for ~3,000 - 5,000 nuclei/uL. Transposition, barcoding, and library preparation were performed according to the 10x Genomics Chromium Next GEM Single Cell Multiome protocol CG000338 Rev E.