Illumina HiSeq 2000 paired end sequencing; RNA-seq of Pdx1+ mouse Islet during development
RNA-seq of Pdx1+ mouse Islet during development
Isolated islets were GFP sorted using mouse lines that expressed GFP with a minimal Pdx1 promoter. No MARIS sorting. After sorting, cells were pelleted by centrifugation at 3000 g for 5min at 4C. The supernatant was discarded. Total RNA was isolated from the pellet using the RecoverAll Total Nucleic Acid Isolation kit (Ambion), starting at the protease digestion stage of manufacturer-recommended protocol. The following modification to the isolation procedure was made: instead of incubating cells in digestion buffer for 15 minutes at 50C and 15 minutes at 80C, we carried out the incubation for 3 hours at 50C. Cell lysates were frozen at -80C overnight before continuing the RNA isolation by the manufacturer's instructions. NA
Experimental Factor: developmental stage
adult