Time-lapse RNA sequencing during gastruloid development at different concentrations of retinol
Time-lapse RNA sequencing during gastruloid development at different concentrations of retinol
Gastruloids were generated from Rarb2-LacZ mESCs reporter line with 0 nM, 300 nM, 600nM and 1200 nM retinol. E14 Rarb2-LacZ mESCs and 24 h, 48 h, 72 h, 96 h, and 120 h after aggregation; gastruloids (approximately n= 40-48 and N=3) were collected, washed with ice- cold PBS three times and 500 Âµl TRIzol (Thermo Fisher Scientific,15596018) was added. 2 or 3 replicates were generated for each condition. After TRIzol addition, samples were dissociated using a p1000 pipette and stored at -80oC until RNA isolation. For RNA isolation, the samples were incubated at room temperature until defrosted. Standard TRIzol extraction was performed.  RNA extraction and cDNA library preparation were performed according to the SORTseq protocol using specific CELseq2 barcodes and TruSeq small RNA adaptors. Metadata is provided in the metadata_samples.tsv file.