GSM7767327: Kidney Dehydrated rep2; Rattus norvegicus; RNA-Seq
Male Sprague-Dawley rats were killed by striking the cranium followed by decapitation between 10am and 12pm. Whole kidneys were dissected from each animal and placed on dry ice. Frozen tissue was added to a pre-cooled mortar containing a small volume of liquid nitrogen. The tissue was then ground to a fine powder using a pestle and the nitrogen allowed to evaporate. The powdered tissue was aliquoted, weighed, and stored at -80◦C. Cells were lysed by adding no more than 100mg/ml of Trizol (Qiazol 79306, Qiagen) to each powdered tissue sample, then vortexed for 2 minutes, then incubated on ice for 10 minutes. Lysed samples were spun at 12000xg for 10 minutes at 4◦C and the supernatant was transferred to a new tube. 1/5 volume of Chloroform (99.9%, 22711.244, VWR) was added to each sample and vortexed for 15 seconds until the solution looks like strawberry milkshake. Samples were then spun at 12000xg for 15 minutes at 4◦C. 250µl of the top RNA-containing aqueous phase was removed without contamination from the white interphase. RNA was extracted using Zymo Direct-Zol RNA miniprep (R2052, Zymo) as per manufacturer's instructions.