GSM8780562: K562, DMSO treated, Replicate 1, guideRNA library; Homo sapiens; OTHER
Adherent cells (iPSCs) were dissociated into single cell suspension using accutase and then counted using the automated cell counter Countess. Suspension cells (K562) were directly counted using countess. Appropriate number of cells were used for nuclear extraction and tagmentation following the 10x Multiome protocol. The nuclei were then encapsulated in Gel-bead in Emulsion (GEMs) using the 10X chromium controller and Chromium Next GEM Single Cell Multiome ATAC Kit A. RNA and ATAC libraries were constructed according to 10X instructions. For generating guide RNA libraries during the GEM Generation and step, a duplex splint oligo was spiked into the barcoding reaction mix to allow reverse transcription of gRNAs and ligation onto the capture oligo of 10X multiome gel beads. The resulting cDNA from captured guide RNAs is amplified to make an Illumina-compatible sequencing library containing P5 and P7 adaptors as well as sample indexes.