GSM8775405: E16.5_Wt1creER_Myrf_tdTom_mutant_rep1; Mus musculus; RNA-Seq
For Single Cell RNA-Sequencing, timed Myrf Fl/Fl;R26TdTomato/R26TdTomato x Wt1creERT2;Myrf Fl/Fl mating's were set up and pregnant females were injected with Tamoxifen I.P at E12.5. Females were euthanized at E16.5 and mutant and Cre+ embryos were screened for TdTom+ fluorescence. Mutant and Cre Control lungs were dissected and genotyped based on visual lung/cell shape phenotypes. All genotypes were later confirmed with PCR. Mutant and Cre Control lungs were pooled (3-5 per genotype) and lungs were dissociated into single cells. Samples were sorted using the BD FACSAria II (BD Biosciences) and gating was set up using single color controls. Approximately 50,000 TdTom+Dapi- Mutant and Controls were collected. Approximately 16,000 cells for each genotype were loaded into the 10x Chromium Controller (10x Genomics). libraries were constructed using manufacturers protocol (Chromium Single Cell 3' V3 Kit,10x Genomics)