RNA Purification from Nucleated Blood:
- Add 200 µl 1X DNA/RNA Protection Reagent to 10 µl nucleated blood. Mix by pipetting.
- Add 160 µl Proteinase K Reaction Buffer and 4 µl Monarch Proteinase K. Mix well.
- Incubate at 55°C for 30 min (Note: Optimal incubation times may vary).
- Add an equal volume of isopropanol and mix by vortexing.
- Transfer the sample to a gDNA Removal Column fitted with a collection tube and spin for 30 seconds; discard the flow-through. Transfer column to a new collection tube.
- Add 200 µl Monarch RNA Lysis Buffer to the matrix and let stand for 5 min. Spin for 30 seconds. SAVE THE FLOW-THROUGH and discard the gDNA Removal Column.
- Add an equal volume of ethanol to the flow-through and mix thoroughly by pipetting.
- Proceed to Step 4 of Part 2: RNA Binding and Elution in the Product Manual (Loading sample onto the RNA Purification Column).