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Invitrogen™ RNAlater™ Stabilization Solution

Product Code. 10427114
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Product Code. 10427114 Supplier Invitrogen™ Supplier No. AM7021

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Description

RNAlater RNA 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.

RNAlater RNA 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. Tissue pieces can be harvested and submerged in RNAlater RNA Stabilization Solution for storage without jeopardizing the quality or quantity of RNA obtained after subsequent RNA isolation. Advantages of using RNAlater RNA Stabilization Solution:

• Effectiveness—stabilize RNA for 1 day at 37°C, 1 week at 25°C, 1 month at 4°C, or indefinitely at -20°C
• Simplicity—a single reagent that immediately inactivates RNases and stabilizes RNA within tissues or cells
• Convenience—no need to freeze samples in liquid nitrogen or rush samples back to the lab freezer
• Mobility—perfect for tissue collection 'in the field'
• Versatility—compatible with many RNA isolation procedures, including most RNA isolation kits

Applications
RNAlater RNA Stabilization Solution has been tested on a variety of mammalian tissues, plants, E. coli, Xenopus, fish, and Drosophila. It is ideal for:
• Protecting RNA integrity in tissues rich in RNases
• Collecting samples from different time points without having to process the samples from each time point immediately
• Archiving tissues for future microdissection
• Submerging animal cavities or organs to stabilize RNA during lengthy dissection procedures
• Collecting samples at locations (e.g., hospitals, field sites, the space shuttle) where immediate RNA isolation is not possible
• Shipping samples on wet ice or even at room temperature if shipped overnight

RNAlater RNA Stabilization Solution procedure
The dissected tissue (less than 0.5 cm in any one dimension) is simply submerged in approximately 5 volumes of RNAlater solution at room temperature. The solution permeates the cells, stabilizing the RNA. The sample can then be stored indefinitely at -20°C (the tissue does not freeze), at 4°C for up to a month, or at 25°C for up to a week. For RNA isolation, the tissue is simply removed from RNAlater solution and treated as though it had just been harvested. Most tissues can be transferred directly to a lysis buffer and homogenized. Samples treated with RNAlater solution and then frozen can be ground with mortar and pestle or thawed and processed like fresh tissue without concern for cell rupture and release of RNases since the RNases have already been inactivated. Cells can be spun out and then added to lysis buffer, or in some cases, RNAlater solution can be added along with the cells directly to the lysis buffer.

Compatible with a variety of procedures
RNAlater RNA Stabilization Solution is compatible with one-step RNA isolation methods, such as TRIzol Reagent; with glass binding methods such as Qiagen's RNeasy or the Ambion RNAqueous kit; with acid phenol extraction methods such as the Ambion ToTALLY RNA kit; and with methods that use oligo(dT) selection of mRNA, such as the Ambion Poly(A)Purist™ kit. In-house research, as well recently published independent research, indicates that the use of RNAlater RNA Stabilization Solution for tissue storage does not affect the outcome of subsequent RNA expression analysis experiments compared to other processing methods.

Order Info

Shipping Condition: Room temperature

TRUSTED_SUSTAINABILITY

Specifications

Content And Storage 500 mL RNAlater Stabilization Solution; room temperature
Description RNA Storage Reagent
Sample Type Bacteria, Cultured Cells, Tissue, Yeast
Quantity 500 mL
Shipping Condition Room Temperature

Frequently Asked Questions (FAQs)

I see a precipitate in my RNAlater Stabilization Solution. What should I do?

Heat the solution to 37 degrees C for 15 minutes and agitate to redissolve it.
What can I use to protect RNA in my frozen tissue sample?

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.
Can RNAlater Stabilization Solution be used to preserve samples for laser capture microdissection (LCM)?

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.
Have you tested temperature stability of RNA stored in RNAlater Stabilization Solution?

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.
Can cells treated with RNAlater Stabilization Solution be FACS sorted? How about mass spectrometry?

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.
Can tissue be homogenized immediately after immersion in RNAlater Stabilization Solution?

No, the tissue in RNAlater Stabilization Solution must be stored at 4 degrees C overnight.
Can RNAlater Stabilization Solution be used with gram-positive bacteria?

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.
Does RNAlater Stabilization Solution inactivate bacteria?

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.
Does RNAlater Stabilization Solution inactivate viruses?

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.

How long will an RNA population remain stable and unchanged in RNAlater Stabilization Solution?

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.
Can DNA be isolated from samples stored in RNAlater Stabilization Solution? How about protein?

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.

I'm about to lyse my tissue or cell samples that are stored in RNAlater Stabilization Solution. Do you have any suggestions of how to do this?

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).

How do I use RNAlater Stabilization Solution?

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.

Is RNAlater Stabilization Solution compatible with my system?

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.
What can I use on my cells/tissues to protect the RNA?

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.
With the direct lysis approach to DNA/RNA analysis, can I use cells that are frozen or stabilized with RNAlater solution?

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.
Are MagMax kits compatible with RNAlater reagent-treated samples?

Only MagMax96 for Microarrays Total RNA Isolation Kit (Cat. No. AM1839) is compatible, since it uses Trizol reagent and most of the salt carry-over from RNAlater reagent is eliminated during the lysis steps and partitioned from the aqueous RNA layer on top. For other MagMax kits, optimization is needed. High salt carry over is not optimal for MagMax RNA isolation steps.
Can I use samples treated with RNAlater reagent for protein analysis?

RNAlater reagent will denature proteins; therefore, protein obtained from samples stored in RNAlater reagent are suitable for applications such as Western blotting or gel electrophoresis, but not for applications that require native protein.
Is it possible to reuse RNAlater reagent and RNAlater ICE reagent?

We do not recommend reusing RNAlater reagent or RNAlater ICE reagent.
Can RNAlater reagent be used for liquid samples?

RNAlater reagent is not suitable for liquid samples since it is already at a 1X concentration. When customers must absolutely preserve their liquid samples with RNAlater reagent, the only recommendation we have is to add 15 to 20 volumes of this solution to one volume of their sample. For example, if you have 1 mL of a liquid sample, you would add a minimum of 15 mL of RNAlater reagent to the sample to account for the dilution effect. So a 30 mL sample would require at least 450 mL of RNAlater reagent to minimize the effect of diluting the RNAlater reagent and maintain preservation of the RNA.
I saw some precipitates in my RNAlater bottle. Is it normal? Can I still use it?

RNAlater reagent is a high-salt solution that is close to its saturation point. If RNAlater reagent has crystals or precipitates, heat at 37 degrees C with agitation, redissolve the crystals, and use as directed in the manual. If you have tissues in RNAlater reagent and find that there are crystals, the tissue should be moved to a fresh tube, taking care to leave the crystals behind. For cells, it is best to resuspend the cells and allow the crystals to settle to the bottom. Next, the cells resuspended in RNAlater reagent should be spun and the RNAlater reagent removed. The cell pellet can be stored by itself without RNAlater reagent at -20 degrees C or -80 degrees C, as long as the cells have been incubated in RNAlater reagent overnight.
Are RNAlater Stabilization Solution and RNAlater-ICE Frozen Tissue Transition Solution chemically reactive with oxidizing agents?

RNAlater Stabilization Solution and RNAlater-ICE Frozen Transition Solution are known to react with hypochlorite solutions, such as common bleach. The reaction releases chlorine gas and generates heat. A similar reaction may occur with other oxidizing agents. If you suspect that samples may contain bleach or other oxidizing reagents, we recommend working in a fume hood with adequate protective clothing and equipment.
Can I add RNAlater reagent to samples that are already frozen?

A different product, RNAlater-ICE reagent, is used with samples that are already frozen. RNAlater-ICE reagent transitions tissue from a frozen to a non-frozen state. The frozen tissue is simply placed in RNAlater-ICE reagent and left at -20 degrees C overnight. Treated tissues can then be used directly in standard homogenization and isolation protocols and processed like fresh tissue.
RNAlater reagent and RNAlater-ICE reagent provide flexibility for sample collection and storage, and help ensure that high quality RNA is preserved in samples. Both are available in a variety of convenient sizes.
How can I ship my sample in RNAlater reagent?

Samples in RNAlater reagent can safely be shipped on wet ice for several days. For longer shipping times use dry ice.
Which downstream applications can be used with tissue stored in RNAlater reagent?

Tissue stored in RNAlater reagent is compatible with all commonly used RNA isolation methods, including single reagent isolation products like TRIzol reagent, and all of our Ambion RNA isolation kits. It is also possible to extract both genomic DNA and total protein from samples stored in RNAlater reagent. RNAlater reagent will denature proteins, so it is only compatible with routine protein analyses such as western blotting and 2D gel electrophoresis that do not require native protein.
Has RNAlater reagent been tested with my tissue or with blood samples?

RNAlater reagent has been successfully tested with many different tissues, (including brain, heart, kidney, liver, skeletal muscle, fat, lung) and cell types (E. coli, Drosophila, cultured mammalian cells, and some plant cells). RNAlater reagent can also be used to store anticoagulated whole blood or the white blood cell fraction of whole blood. Ambion RiboPure-Blood RNA Isolation Kit incorporates RNAlater reagent and provides instructions on how best to use RNAlater reagent with blood.
Does storing samples in RNAlater reagent have advantages over freezing samples on dry ice or in liquid nitrogen?

Processing samples that were stored in RNAlater reagent is much easier than using frozen samples. Frozen samples must be ground to a powder and then the frozen powder must be transferred to a tube for homogenization. This procedure is laborious, messy, risks loss of sample, and perhaps most importantly, may lead to sample thawing, which can compromise RNA integrity. Samples stored in RNAlater reagent are protected from RNases as long as the tissue remains in the solution, and they can typically be disrupted using the simpler methods appropriate for freshly collected samples (grinding in liquid nitrogen is only required for extremely hard or tough tissues such as bone or tumor tissue). Thus, in addition to making sample disruption easier, storage in RNAlater reagent eliminates the risks of sample loss and mess due to transferring or thawing frozen powdered tissue.
How do I use RNAlater reagent to store my tissue/cell sample?

For fresh tissue, simply cut samples to a maximum thickness of 0.5 cm in any one dimension and submerge in 5 volumes of RNAlater reagent.

Cultured cells should be pelleted, resuspended in a small volume of PBS, then mixed with 5-10 volumes of RNAlater reagent.

Once in RNAlater reagent, samples can be stored for up to 1 day at 37 degrees C, 1 week at 25 degrees C, 1 month or more at 4 degrees C, and long-term at -20 degrees C or -80 degrees C.
Can I use RNAlater Stabilization Solution for preservation of tissue with freezing before using it for RNA isolation?

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).
Can I use RNAlater Stabilization Solution for long-term preservation of membrane structure (membrane of cell, nuclei, organelle, vesicle, etc.) in tissue with no freezing?

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
Will RNAlater Stabilization Solution freeze at -80 degrees C?

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
Is RNAlater Stabilization Solution a tissue preservation solution similar to a histology fixative (i.e., formalin, glutaraldehyde)?

No, RNAlater Stabilization Solution does not involve chemical fixation or crosslinking. It works by denaturing proteins, including RNase.
How can I spin cells out of RNAlater Stabilization Solution?

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.
Can I isolate genomic DNA from tissue preserved in RNAlater Stabilization Solution (Cat. No. AM7024)?

Yes, you can isolate genomic DNA from tissue preserved in RNAlater Stabilization Solution (Cat. No. AM7024).
Do you offer a sterile version of RNAlater Stabilization Solution?

Unfortunately, we do not offer a sterile version of RNAlater Stabilization Solution.
I am having trouble performing PCR amplification after DNA isolation with RNAlater-treated samples. Does RNAlater have an impact on downstream PCR applications?

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.
What is the difference between RNAlater Stabilization Solution and RNaseOUT Recombinant Ribonuclease Inhibitor? When do I use one over the other?

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.
Is the MagMAX-96 for Microarrays Total RNA Isolation Kit (Cat. No. AM1839) compatible with samples treated with RNAlater Stabilization Solution (Cat. No. AM7021)?

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.

Do Tempus Blood RNA Tubes (Cat. No. 4342792) stabilize RNA similarly to RNAlater Stabilization Solution (Cat. No. AM7021)?

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.

For Research Use Only. Not for use in diagnostic procedures.

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