Guidelines for Sample Stabilization Using the Monarch StabiLyse™ DNA/RNA Buffer

Monarch StabiLyse DNA/RNA Buffer is a dual function solution that preserves integrity of and effectively lyses cells, enabling seamless sample storage and RNA purification without the need to remove the reagent prior to cell lysis.

General usage guidelines for common sample types are listed below. Certain individual sample types may be successfully preserved for longer times and/or at higher temperatures and researchers are encouraged to empirically determine optimal conditions for their sample of interest.

Sample Stabilization of Cells

For cultured mammalian cells, cell pellets can be preserved by resuspending in 350 µl Monarch StabiLyse DNA/RNA Buffer and stored according to storage duration recommendation below.
This method can also be used for adherent cells grown in tissue culture plates, by using appropriate volume of Monarch StabiLyse DNA/RNA Buffer depending on well volume.
For very low cell inputs, such as 100 cells, when pelleting is not feasible, add Monarch StabiLyse DNA/RNA Buffer directly to the cell suspension making the starting volume 350 µl or 700 µl.
For processing samples stored in Monarch StabiLyse DNA/RNA Buffer, allow samples to equilibrate to room temperature. If needed, make up the volume of cell sample to 350 µl or 700 µl to align with the volume recommendation for the cell number. Pipette mix gently but thoroughly to ensure homogenous lysate. Proceed with Step 1 of Part II: RNA Elution and Binding.

Sample Stabilization of Solid Tissues

Stabilization of solid tissue in Monarch StabiLyse DNA/RNA Buffer can be done at the time of harvest using one of the following approaches:

1. Prepare a 1:1 dilution of Monarch StabiLyse DNA/RNA Buffer with Nuclease-free Water. Harvest the tissue sample by working quickly and submerge the tissue piece in the volume of Monarch StabiLyse DNA/RNA Buffer + Nuclease-free Water described in the Tissue protocol. For example, for 10 mg tissue, submerge in 400 µl of StabiLyse + Water solution (1:1). Store the sample according to storage duration recommendation below. For processing samples stored in Monarch StabiLyse DNA/RNA Buffer, allow samples to equilibrate to room temperature. Pipette mix gently but thoroughly to ensure homogenous lysate. Proceed with mechanical homogenization (Step 2 of Tissue lysis protocol) or with Proteinase K treatment (Step 8 of Tissue lysis protocol).
2. Harvest the tissue sample by working quickly and submerge the tissue piece in the volume of StabiLyse recommended in the first step of the extraction process and store according to the storage recommendations below. For example, for 10 mg tissue, submerge in 200 µl of Monarch StabiLyse DNA/RNA Buffer. When ready for extraction, proceed with addition of equal volume of Nuclease-free Water (Step 3 of Tissue lysis protocol) and continue with the protocol steps.

Storage of Samples During Processing

When working with a large number of samples, users may find it preferrable to process samples in batches to maintain RNA integrity and consistency across sample groups. Homogenized samples in Monarch StabiLyse DNA/RNA Buffer can be frozen and –20°Cor –80°C for processing at a later time. When ready for extractions, thaw and allow all samples to reach room temperature before processing.

Sample Storage Recommendation

At 25°C: up to 1 week
At 4°C: up to 4 weeks
At -20°C or -80°C: 4 weeks or more