title: Single cell RNAseq of haploidized HCT116 colorectal carcinoma cells expressing specific p53 mutations treated with either DMSO or Nutlin
design: Single cell RNAseq of haploidized HCT116 colorectal carcinoma cells expressing specific p53 mutations treated with either DMSO or Nutlin
library_construction_protocol: Ten individually generated single cell clones of different HCT116 mut/Δ cell lines were labeled by lentiviral transduction with a specific combination of expressed SPLINTR barcodes (gift from Mark Dawson, Addgene #179774 and #179775). TP53 variants were encoded by GFP-coupled barcodes, clone IDs by mCherry-coupled barcodes.  All barcoded cell clones were pooled and split into two aliquots. One aliquot was treated with 10 µM N3a, the other with the corresponding volume DMSO for 24 h.   The Evercode WT v2 kit (Parse Biosciences) uses the fixed cells itself as reaction vessel for the reserve transcription so there is no nucleic acid extraction as such. Fixation and permeabilization was performed using the Evercode Cell Fixation v3 product according to manufacturer's instructions. Single-cell RNA sequencing libraries were prepared from both pools using the Evercode WT v2 kit (Parse Biosciences) according to manufacturer's protocol. Quality of sequencing libraries was controlled on a Bioanalyzer using the Agilent High Sensitivity DNA Kit (Agilent) and quantified with the Qubit dsDNA-HS-Assay-Kit (Thermo Fisher Scientific).