A new rapid method for selective extraction of RNA from fixed mammalian tissues.
Treatment of formalin-fixed mammalian tissues with concentrated or 50% phosphoric acid at 5 degrees C for 20 and 50 min. respectively reveals complete extraction of RNA as judged by methyl green followed by staining with pyronin. This procedure also causes depolymerisation of DNA as indicated by the red staining of the nuclei. Sections treated with concentrated phosphoric acid at 5 degrees C for 30 min. causes disruption of the double helical structure of DNA what results in the depression of the pyronin staining. Similarly treated sections show Feulgen positive nuclei. Treatment of sections in 25 % phosphoric acid at 60 degrees C for 15 min. followed by staining with methyl green and pyronin show red nuclei, nucleoli and the cytoplasm. This indicates that extraction of RNA is only possible in cold and not at elevated temperature.
D000818:Animals; D003080:Cold Temperature; D007668:Kidney / Q000032:analysis*; D008099:Liver / Q000032:analysis*; D008722:Methods; D010756:Phosphoric Acids; D011754:Pyronine; D012313:RNA / Q000302:isolation & purification*; D051381:Rats; D013194:Staining and Labeling
D010756:Phosphoric Acids; D012313:RNA; D011754:Pyronine