I decided to run another RNA isolation today with 8 more samples: D03, D04, D13, D14, T03, T04, T13, T14
Protocol was as follows:
- 500 µL of RNAzol RT was added to a clean tube.
- Tissue samples were removed and a small section was cut out for RNA extraction.
- Tissue portions were placed in the tube and an additional 500 µL of RNAzol RT was added to bring the volume up to 1mL.
- The samples were vortexed vigorously for 10 seconds
- Samples were incubated at room temperature for 5 minutes.
- 400 µL of DEPC-water was added to the samples.
- Samples were centrifuged for 15 minutes at 12,000 g.
- 750 µL of the supernatant was transferred to a new, clean tube and an equal volume of isopropanol was added to the sample.
- The samples were vortexed vigorously for 10 seconds.
- Samples were incubated at room temperature for 5 minutes.
- Samples were centrifuged for 15 minutes at 12,000 g.
Due to time constraints, I decided to finish up the extraction later/quantify RNA using the Qubit and stored the RNA pellet suspended in isopropanol in the -80 freezer.