RNAlater™ Stabilization Solution with Manual - FAQs

Heat the solution to 37 degrees C for 15 minutes and agitate to redissolve it.

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RNAlater-ICE Frozen Tissue Transition Solution can be used to submerge a frozen sample, then thaw it overnight at -20 degrees C or colder. Once thawed, tissues can then be processed like fresh tissues using standard RNA isolation procedures.

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While we have not tested this in-house, there is evidence that it is possible to use tissues preserved in RNAlater Stabilization Solution and then perform laser capture microdissection. Please see the reference below:
Thelen P, Burfeind P, Grzmil M et al. (2004) cDNA microarray analysis with amplified RNA after isolation of intact cellular RNA from neoplastic and non-neoplastic prostate tissue separated by laser microdissections. Int J Oncol 24(5):1085-1092.

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RNA stored in RNAlater Stabilization Solution is stable for 1 day at 37 degrees C, 1 week at 25 degrees C, 1 month at 4 degrees C, or long-term at -20 degrees C.

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Yes, you should be able to sort cells by fluorescence-activated cell sorting (FACS) after treatment with RNAlater Stabilization Solution. If you run into problems due to the viscosity of RNAlater Stabilization Solution, you may need to dilute it with cold nuclease-free water. This will not affect the protection of the RNA. Samples should be processed quickly and as cold as possible. Read more about this procedure and view references here (http://www.thermofisher.com/us/en/home/references/Invitrogen-tech-support/rna-isolation/tech-notes/facs-into-rnalater-solution-for-gene-profiling.html). You should also be able to perform mass spectrophotometry on samples treated with RNAlater Stabilization Solution. Please note that salt can form adducts on the protein at the desorption/ionization step. To avoid this problem, dialyze the sample to get rid of all salt.

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No, the tissue in RNAlater Stabilization Solution must be stored at 4 degrees C overnight.

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Yes, RNAlater Stabilization Solution has been used with Staphylococcus epidermidis isolates. Here's a reference describing how researchers used the solution with gram-positive bacteria:

Handke LD, Conlon KM, Slater SR et al. (2004) Genetic and phenotypic analysis of biofilm phenotypic variation in multiple Staphylococcus epidermidis isolates. J Med Microbiol 53(Pt5):367-344.

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RNAlater Stabilization Solution is bacteriostatic. Although bacteria do not grow in RNAlater solution, the cells remain intact. E. coli cells stored in RNAlater Stabilization Solution for 1 month at 4 degrees C are intact and yield undegraded RNA.

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While we have not tested this in house, these references suggest that the virus remains active after treatment with RNAlater Stabilization Solution:

- Uhlenhaut C, Kracht M (2005) Viral infectivity is maintained by an RNA protection buffer. J Virol Methods 128(1-2):189-191.
- Kurth A (2007) Possible biohazard risk from infectious tissue and culture cells preserved with RNAlater. Clin Chem3(7):1389-1390.

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RNA from RNAlater treated tissue was evaluated after -20 degrees C storage for over 2 years, indicating no systematic bias introduced by storage in RNAlater Stabilization Solution. For more information, visit our webpage at http://www.thermofisher.com/us/en/home/references/ambion-tech-support/rna-isolation/tech-notes/rna-remains-stable-during-long-term-tissue-storage.html.

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Yes, DNA can be isolated from samples stored in RNAlater Stabilization Solution. Please view the protocol here: https://www.thermofisher.com/us/en/home/references/protocols/nucleic-acid-purification-and-analysis/rna-protocol/genomic-dna-preparation-from-rnalater-preserved-tissues.html. Proteins will be preserved in RNAlater Stabilization Solution; however, the solution will denature the proteins. Therefore, the protein obtained from samples stored in it will still be suitable for applications such as western blotting or 2D gel electrophoresis, but not for applications that require native protein.

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For your tissue samples, we recommend removing the tissue treated with RNAlater Stabilization Solution from the excess RNAlater solution after incubation overnight at 4 degrees C, blotting the tissue on an absorbent lab wipe or paper towel, and proceeding to lysis.

For your cell samples, pellet the cells, then pour off the excess RNAlater supernatant. Cells are generally less fragile after being stored in RNAlater Stabilization Solution and most cells can be centrifuged at higher speeds without causing cell lysis (5,000 x g).

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Trim the tissue to be less than 0.5 cm in at least one dimension and simply submerge it in 5 volumes of RNAlater Stabilization Solution (e.g., a 0.5 g sample requires about 2.5 mL of RNAlater Stabilization Solution). You can store most samples at 4 degrees C for up to one month, at 25 degrees C for up to one week, or at -20 degrees C indefinitely.

For RNA isolation, simply remove the tissue from RNAlater Stabilization Solution and treat it as though it was just harvested. Most tissues can be homogenized directly in lysis buffer, although harder tissues such as bone may require freezing in liquid nitrogen and grinding.

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RNAlater Stabilization Solution is compatible with standard RNA isolation kits as long as the RNAlater Stabilization Solution is removed prior to the RNA isolation procedure.

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RNAlater Stabilization Solution is an aqueous tissue storage reagent that protects cellular RNA in intact, unfrozen tissue samples. RNAlater Stabilization Solution eliminates the need to immediately process your tissue samples or to freeze them in liquid nitrogen for later processing.

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For the Cells-to-CT lysis reaction, you can use fresh cells, frozen cells, or cells that have been stabilized with RNAlater solution. You just want to ensure that cells were washed once with PBS before going into the lysis reaction.

Yes, for the purpose of RNA preservation in tissue, RNAlater Stabilization Solution will work as well as snap freezing ( https://www.thermofisher.com/us/en/home/references/ambion-tech-support/rna-isolation/tech-notes/rna-remains-stable-during-long-term-tissue-storage.html).

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We have only tested RNAlater Stabilization Solution for preservation of tissue up to 1 month at 4 degrees C before isolation of RNA. Therefore, we cannot recommend using it for long-term preservation of tissue with no freezing

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Yes, RNAlater Stabilization Solution will freeze at -80 degrees C. We recommend treating your tissue overnight at 4 degrees C with RNAlater and then removing RNAlater prior to freezing at -80 degrees C

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No, RNAlater Stabilization Solution does not involve chemical fixation or crosslinking. It works by denaturing proteins, including RNase.

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Dilute the solution 1:1 with cold PBS and centrifuge at up to 5000 x g (start at lower speed and increase until the cells pellet). Transfer the supernatant to a new tube and discard the pellet.

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Yes, you can isolate genomic DNA from tissue preserved in RNAlater Stabilization Solution (Cat. No. AM7024).

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Unfortunately, we do not offer a sterile version of RNAlater Stabilization Solution.

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RNAlater should not impact downstream PCR amplification as long as the sample has been properly cleaned before proceeding with DNA isolation, as stated in the RNAlater Stabilization Solution manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/7020M.pdf) on page 7.

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RNAlater Stabilization Solution stabilizes and protects cellular RNA in intact, unfrozen tissue samples, eliminating the need to immediately process tissue samples or to freeze samples in liquid nitrogen for later processing. In other words, it is an RNA preserving reagent.

RNaseOUT Recombinant Ribonuclease Inhibitor is a potent non-competitive inhibitor of pancreatic-type ribonucleases such as RNase A, and is used to avoid RNA degradation in a variety of applications such as cDNA synthesis, RT-PCR, and in vitro transcription and translation. RNAseOUT inhibits RNase A, RNAse B, and RNAse C. In other words, RNaseOUT Recombinant Ribonuclease Inhibitor is mainly used in an assay to prevent RNA degradation.

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Yes, MagMAX-96 for Microarrays Total RNA Isolation Kit is compatible with the RNAlater solution, because the kit uses Tri-Reagent. Therefore, most of the salts are removed during the lysis steps and partitioned from aqueous RNA layer on top. Additionally, salt carry-over from the RNAlater solution is not an issue.

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Both Tempus Blood RNA Tubes and RNAlater Stabilization Solution employ the same effective stabilization of mRNA expression profiles, and eliminate the need to isolate the RNA immediately after sample collection. However, there are some key differences:
- Components in each stabilization solution are different. The exact components and differences are proprietary.
- The RNAlater Stabilization Solution preserves RNA from degradation without lysing blood cells.
- Samples in RNAlater Stabilization Solution can be safely stored for up to 1 week at 25 degrees C, or up to 1 month at 4 degrees C.

Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.

Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.