FAQ: Can I lyse my adherent cells directly in their wells when using the Monarch Total RNA Miniprep Kit?

Yes. Our standard protocol for cultured mammalian cells can be modified to lyse cells directly in the wells of a multiwell plate.

For immediate processing:

1. Remove the media from the well and rinse with PBS.
2. Add Monarch RNA Lysis Buffer (NEB #T2012) directly to the well according to the table below:

   Multiwell plate size	Volume of Monarch RNA Lysis Buffer
   6 well	> 600 μl
   12 well	300 - 600 μl
   24 well	300 μl
   48 well	300 μl

3. Mix thoroughly and proceed to Step 1 of Part 2: RNA Binding and Elution.

 For storage:

1. Remove the media from the wells and rinse with PBS.
2. Add 1X Monarch DNA/RNA Protection Reagent (NEB #T2011) (be sure to dilute the 2X concentrate) directly to the well according to the table below:

   Multiwell plate size	Volume of 1X Monarch DNA/RNA Protection Reagent
   6 well	>600μl
   12 well	300 - 600μl
   24 well	300μl
   48 well	300μl

3. Mix thoroughly and store samples until ready for processing.  Samples may be frozen for long-term storage or may be refrigerated or maintained at room temperature for medium-term or short-term storage (see Sample Stabilization and Storage Instructions or refer to the product manual for more details).
4. When ready to process, add an equal volume of Monarch RNA Lysis Buffer (NEB #T2012) and proceed to Step 1 of Part 2: RNA Binding and Elution.