Guidelines for RNA Purification from Leukocytes (WBCs, PBMCs)

1. The Monarch Total RNA Miniprep Kit (NEB #T2010) can be used to purify up to 1x10⁶ Leukocytes (WBCs, PBMCs).
   
   
2. Be sure to use a 1X solution of Monarch DNA/RNA Protection Reagent. A 1X solution is prepared by diluting the necessary volume of the supplied 2X concentrate with an equal volume of nuclease-free water.
   
   
3. For samples stored in Monarch DNA/RNA Protection Reagent, allow samples to equilibrate to room temperature prior to processing. Ensure the volume of Protection Reagent is consistent with protocol recommendations and adjust if necessary.
   
   
4. Perform the Proteinase K digestion for 30 minutes at 55°C to ensure complete digestion.
   
   
5. Addition of Monarch RNA Lysis Buffer (NEB #T2012) and all subsequent steps should be performed at room temperature.

Additional Resources:

• General Guidelines for Successful RNA Purification Using the Monarch Total RNA Miniprep Kit
  
  
• Guidance on Choosing Sample Input Amounts when Using the Monarch Total RNA Miniprep Kit
  
  
• Troubleshooting Guide for RNA Purification