GSM8990893: TAPseq, MEF cells, gRNA enrichment from GEX, biol rep 1; Mus musculus; RNA-Seq
CROP-RFP-positive cells were flow sorted, fixed in 0.1% formaldehyde for 5 minutes, permeabilized in 0.1% NP-40 for 5 minutes and resuspended in 10X Genomics Diluted Nuclei Buffer at 3x10e6 cells per ml. 5ul (15,000 cells) of fixed/permeabilized cells was input to the 10X library preparation step. GEX & ATAC libraries were generated on the 10X Chromium Controller using the 10X Genomics Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Kit according to the manufacturer's instructions gRNA enrichment was performed on the GEX cDNA library using the TAPseq protocol (Schraivogel et al, 2020, https://doi.org/10.1038/s41592-020-0837-5) mtDNA enrichment was performed on the ATAC library by hybridization enrichment with a bespoke panel of mtDNA probes (IDT) using the IDT xGen Hybridization and Wash Kit according to manufacturer's instructions