GSM3578982: scRNAseq in EB+48hrDox(iAscl1-v5 or iNeurog2 Tubb3::GFP); Mus musculus; RNA-Seq
Single-cell RNA-seq: Cells (iAscl1-v5 and iNeurog2 Tubb3::GFP) were collected 48 hours after Dox induction and washes were done in 1X PBS with 0.04 mg/ml BSA (Thermo Fisher Sci AM2616). Cells were strained with CellTrics 30 µM (Cat #04-004-2326) to remove cell clumps. Equal number of iA and iN Tubb3::GFP cells were pooled to have 1000 cells/ul. 10X Genomics Chromium Single Cell 3' library kit was used to generate single cell library for a targeted cell recovery rate of 10.000 cells (120262 Chromium i7 Multiplex Kit, 120236 Chromium Single Cell 3' Chip Kit v2, 120237 Chromium Single Cell 3' Library & Gel Bead Kit v2). Fragment length distribution of the library was determined by Agilent High Sensitivity DNA D1000 Screentape (5067- 5585) system and the final quantification of the library before pooling was done with KAPA library amplification kit (Roche Lightcycler 480). The libraries were sequenced on Illumina NextSeq 500 High Output using V2.5 chemistry with 26x98 bp - 150 cycles run confirmation at the genomics core facility at NYU.
GEO Accession
GSM3578982