cbil.upenn.edu:RAD.Protocol:2324:Protocol - cbil.upenn.edu:RAD.Protocol:2324:Protocol
Tissue was extracted, immediately placed in GIT Buffer (5 M guanidinium isothiocyanate, 50 mM Tris-HCl pH 7.5, 10 mM EDTA pH 8.0, and 5% 2-mercaptoethanol), and homogenized with a Polytron. Tissue samples were then layered onver 4 ml of 5.7M CsCl then spun at 27,000 RPM overnight. The supernatant was aspirated and the RNA pellet resuspended in DEPC water. Samples were phenol extracted and Ethanol precipitated. The final RNA pellet was dried under vacuum and resuspended in DEPC water.