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Invitrogen™ RNAlater™-ICE Frozen Tissue Transition Solution

Catalog No. AM7030
Encompass
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25 mL
10 x 25 mL
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RNAlater-ICE is a novel reagent for transitioning frozen tissue to a state that is easily processed for extraction of high quality RNA.

RNAlater-ICE is a novel reagent for transitioning frozen tissue to a state that is easily processed for extraction of high quality RNA. Frozen tissues are simply submerged in RNAlater-ICE and allowed to thaw overnight at -20°C. Once thawed, the tissues can be processed like fresh tissues using standard RNA isolation procedures. No more laborious grinding of frozen tissue to safeguard the RNA.

• Process previously frozen tissues like freshly harvested samples
• Thawing tissues in RNAlater-ICE protects RNA from degradation
• No more tissue pulverizing with mortar and pestle and awkward transfer of powder to tube
• Easily apportion frozen tissue samples for multiple experiments
• Quick Freezing Tissues Preserves RNA

Often, tissues that need to be stored prior to RNA isolation are “snap” or “flash” frozen on dry ice or in liquid nitrogen to preserve RNA integrity. RNA in tissue is stable while frozen at -80°C but thawing the tissue prior to or during its disruption can result in RNA degradation. This is true even if the tissue thaws while in the denaturation solution.

Processing Frozen Tissues is Problematic
Frozen tissues are typically ground with a chilled mortar and pestle in order to maintain RNA quality. Liquid nitrogen must be added to the mortar to keep the sample frozen while it is ground. For multiple samples this process is laborious. Either a separate mortar and pestle set is needed for each sample, or the set must be thawed and cleaned after each sample is processed to avoid cross-contamination. Powdered tissue can also thaw during transfer to a homogenization vessel. This often results in formation of clumps that do not readily disperse in the lysis solution, resulting in RNA degradation and loss. Very small samples should be homogenized immediately in lysis solution, which can again be cumbersome if there are multiple samples to process.

Process Frozen Tissue Without Jeopardizing RNA Integrity
RNAlater-ICE solves all of these problems. Simply submerge frozen tissue samples in 10 volumes of RNAlater-ICE and store overnight at -20°C (the solution will remain liquid at these temperatures). As the tissue thaws, RNA integrity is protected. Once treated, tissue can be safely stored at 4°C or even at room temperature (for a limited period of time) and can be further dissected or processed prior to homogenization in a standard RNA isolation lysis buffer. Thus the same frozen tissue sample can be used multiple times for different experiments without compromising RNA integrity.

Specifications

Content And Storage 25 mL RNAlater-ICE Solution; room temperature

Description RNA Storage Reagent
Sample Type Tissue (Frozen)
Quantity 25 mL
Shipping Condition Room Temperature
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.

Is it possible to reuse RNAlater reagent and RNAlater ICE reagent?

We do not recommend reusing RNAlater reagent or RNAlater ICE reagent.

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.

I want to extract cytoplasmic RNA for RNA sequencing from frozen tissues without affecting integrity of the nucleus. Can I homogenize my sample in RNAlater-ICE Frozen Tissue Transition Solution and extract cytoplasmic RNA with an RNA purification kit?

RNAlater-ICE does not lyse the cells and the nucleus should stay intact. The reagent works primarily through protein denaturation. With proper cytoplasmic and nuclear fraction separation, you should be able to isolate the cytoplasmic RNA.


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