GSM2667103: AG1neg_51, Treated single cell RNA-seq; Mus musculus; RNA-Seq
Wild type and K14ΔNβ-cateninER keratinocytes were cultured on 12 well plates in a ratio of 9:1 for a total of 200,000 cells per well and allowed to attach for 24 hours. Subsequently, cells were treated with 4-OHT (200nM) or DMSO as a control. After 24 hours of treatment cells were trypsinized and resuspended as a single cell suspension. Single keratinocytes were captured on a medium-sized (10-17μm) microfluidic chip (C1, Fluidigm). Cells were assessed for viability (LIVE/DEAD assay, Life Technologies) and C1 capture sites were imaged by phase contrast to determine empty and doublet capture sites. Cells were loaded onto the chip at a concentration of 300 cells μl-1. Doublet or non-viable cells were excluded from later analysis. Cell lysis, reverse transcription, and cDNA amplification were performed on the C1 Single-Cell Auto Prep IFC, as per the manufacturer’s instructions. For cDNA synthesis the SMART-Seq v4 Ultra Low Input RNA Kit (Clontech) was used. Single cell Illumina NGS libraries were constructed with Nextera XT DNA Sample Prep kit (Illumina). Sequencing was performed on Illumina HiSeq4000 (Illumina) using 100bp paired-end reads.
GEO
GSM2667103
GEO Accession
GSM2667103