Home FAQs Can the Monarch Total RNA Miniprep Kit (NEB #T2010) be used to purify RNA from nucleated blood (e.g. bird, fish, reptile)?

FAQ: Can the Monarch Total RNA Miniprep Kit (NEB #T2010) be used to purify RNA from nucleated blood (e.g. bird, fish, reptile)?

Yes. We recommend stabilizing nucleated blood in Monarch DNA/RNA Protection Reagent (NEB #T2011) and processing samples as described below. 
 
RNA Purification from Nucleated Blood:
  1. Add 200 µl 1X DNA/RNA Protection Reagent to 10 µl nucleated blood. Mix by pipetting.
  2. Add 160 µl Proteinase K Reaction Buffer and 4 µl Monarch Proteinase K.  Mix well.
  3. Incubate at 55°C for 30 min (Note: Optimal incubation times may vary).
  4. Add an equal volume of isopropanol and mix by vortexing.
  5. 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.
  6. 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.
  7. Add an equal volume of ethanol to the flow-through and mix thoroughly by pipetting.
  8. Proceed to Step 4 of Part 2: RNA Binding and Elution in the Product Manual (Loading sample onto the RNA Purification Column).