RNAstable^® Protocol

Product Nos. 93220-001, 93221-001, and 90220-001

Product Description

RNAstable^® is a unique storage medium that preserves RNA samples at room temperature. RNAstable^® enables you to store RNA samples for months. Using RNAstable^®, you can completely recover your RNA samples by simple rehydration - just add water. Rehydrated samples are ready for immediate use, without the need for further purification.

Each tube or plate contains RNAstable^® provided as a coating at the bottom of the tube or well, which protects picogram to microgram amounts of RNA.

Precautions and Disclaimer

This product is for R&D use only, not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices.

Storage/Stability

RNAstable^® products and kits should be stored dry in their original unopened packaging at ambient laboratory temperatures ready for use. The RNAstable^® products are supplied in moisture-barrier foil bags along with a desiccant pack. The sample pouch allows for dry storage, even in uncontrolled humidity environments, and serves as an alternative to storage in a dry storage cabinet.

Procedures

Notes on Sample Preparation

Types of RNA. Suitable types of RNA that can be stored in RNAstable^® include purified total RNA from cells or tissues, and poly(A) mRNA.

Purification Techniques. Most standard molecular biology techniques and/or commercially available kits are compatible with RNAstable^® storage. Purified RNA that is RNase-free should be resuspended in DEPC- treated water prior to application into RNAstable^®.

Determining yield. The concentration of the RNA sample should be determined prior to sample application into RNAstable^®. Although not essential, applying a known amount of RNA into RNAstable^® for storage can facilitate sample retrieval and subsequent applications. For optimal protection in RNAstable^®, do not apply more than a total of

100 μg of RNA per well or tube in a maximum volume of 100 μL water.

Sample Drying and Storage

1. Determine the amount of purified RNA in the sample and calculate the amount to be applied into RNAstable^® wells or tubes. RNA should be resuspended in DEPC-treated water prior to application on RNAstable^®.
2. Gently add up to 100 μL of the RNA sample directly into the center of each tube or well containing RNAstable^®.
   For optimal sample protection and recovery, the final volume of the sample applied to each well should be ≤20 μL for airdrying; do not exceed applying 100 μL water per tube or well.
3. Mix the sample thoroughly with gentle pipetting (avoid forming air bubbles).
4. Dry the uncovered sample. For convenient air-drying of volumes ≤20 μL, leave the tube or 96-well plate open overnight in laminar flow hood. For volumes ≥20 μL , or for accelerated drying of all, use a vacuum concentrator, without heat (see Table 1 below for drying times).

Table 1 drying times with a vacuum concentrator

Sample Volume (μL)	Drying Times (hrs) Tubes
10–20	0.5
20–30	1.0
30–100	1.5

5. Store samples with a desiccant packet in the original pouch and heat-sealed. Alternatively, store dried samples in a dry storage cabinet at room temperature. RNAstable^® is sensitive to temperature and humidity, therefore it is critical to maintain stored samples at room temperature (15-25 °C) in an environment protected from moisture. The recommended humidity level is ≤ 30% relative humidity.

Sample Recovery

RNA stored in RNAstable^® can be recovered by the addition of water or aqueous buffer. Samples are ready for downstream applications without the need for further purification.

1. Add 10-100 μL of DEPC-treated water or aqueous buffer directly to the dried sample in RNAstable^®.
2. Incubate at room temperature for 15 min to allow complete rehydration.
3. Mix the sample by gently pipetting up and down. Avoid forming bubbles. The rehydrated sample is now ready for use directly in downstream applications.
4. Store unused rehydrated samples at 4 °C or room temperature for up to 3 days. Samples can also be stored frozen
   (-20 °C or -80 °C) for longer time periods. Rehydrated samples in water containing RNAstable^® can be re-dried without loss of efficient sample stabilization. Repeating this process more than 3 times is not recommended.