Total RNA Extraction - East Carolina University：总RNA提取-东卡罗来纳大学

Total RNA ExtractionReagent required: Clorofoam Isopropyl Alcohol 75% Ethanol RNase free water or 5%

Total RNA Extraction 1of 3 Total RNA Extraction Reagent required: Clorofoam Isopropyl Alcohol 75% Ethanol RNase free water or 5% SDS solution Wear Gloves and Lab Coat Safety: Preparations: 1. Use 1M NaOH to clean probe for 15 min., screw back in rinsing with ethanol and water drying it completely. 2. Use 70% ethanol to clean __ntrifuge and repla__ silver rotor with black rotor (located in RNA Zone). 3. Set __ntrifuge to 5˚C at 11500 rpm for 15 min. (see step #9) Pro__dure: 1. Label micro__ntrifuge tubes (tubes at RNA Zone). 2. Obtain tissue samples and pla__ into tubes (embryos to 0.1 __rks, if older then to 0.5 __rks, unsure ASK INSTRUCTOR) 3. Draw out ex__ss Hank’s solution by pipetting 10 μl 4. Add 1000 μl (1ml) TRIzon Reagent 5. Sonicated 15 __all short shocks or til dissolved 6. Incubate at room temperature for 5min. 7. Add 200 μl (0.2ml) chlorofame, then tightly close cap and shake vigorously with hand for 15 sec. Revision 12-14-06