Perturb-seq of a TF library in keratinocytes, using 10x SPLiT-seq.
Perturb-seq using the SPLiT-seq method; this file combines the Cas9 guide RNA and gene expression reads. Primary keratinocytes were differentiated for four days via calcium and high density, then harvested with trypsin, formaldehyde fixed, and frozen with DMSO. After thawing, two rounds of SPLiT-seq were performed: cells were successively barcoded across three oligo plates by reverse transcription (24 wells) and two ligations (96 wells), split into 14-19 sublibraries, lysed, RNA purified with a biotin handle, a template switch reaction performed to add an adapter, and finally successive rounds of PCR to finish the library.