GSM3938455: Control Embryonic Day 18.5 rep3; Mus musculus; RNA-Seq
The dorsal pancreas was dissected from 3 control and 3 Dpy30∆P E18.5 embryos directly into 1 mL of TRIzol. RNA was extracted with 200 uL of chloroform, followed by a combination of 500 uL of 100% isopropanol and 2 uL of glycogen (20 mg/mL). RNA samples were centrifuged at 12,000xg for 5 minutes at 4C and washed with 80% ethanol. The final RNA pellet was air-dried and resuspended in 12 uL of water. RNA was treated with TURBO DNase I (Thermo Fisher Scientific) for 20 minutes at 37C to remove genomic DNA. For each sample, 100 ng of RNA was ribodepleted with the RiboGone kit (Takara Bio). cDNA libraries were prepared from <10 ng RNA using the SMARTer Stranded RNA-seq kit (Clontech Laboratories, Inc.) and SPRIselect beads (Beckman Coulter).