GSM6560658: DMSO 16h rep3; Mus musculus; RNA-Seq
After 7 days of treatment with 600nM 4-Hydroxytamoxifen, organoids were lysed and RNA was extracted following the Trizol RNA isolation protocol (W.M. Keck Foundation Biotechnology Microarryay Resource Laboratory at Yale University) until the end of the phase-separation step. Total RNA cleanup with DNase digestion was performed by addition of 1.5 volumes of absolute ethanol on top of the aqueous phase obtained after the phase-separation and following the Qiagen RNeasy protocol (W.M. Keck Foundation Biotechnology Microarryay Resource Laboratory at Yale University). RNA integrity was analysed on Agilent 2100 bioanalyzer. All conditions were prepared as biological triplicates and sent to BGI Tech Solutions (HongKong)- Co for library preparation and RNA sequencing. Purification of mRNA from total RNA was achieved using oligo(dT)-attached magnetic beads and then fragmented for random hexamer-primed reverse transcription, followed by a second-strand cDNA synthesis.