- Simplifies sample collection—one reagent immediately inactivates RNases and stabilizes RNA within tissues or cells
- More flexibility—no need to freeze samples in liquid nitrogen or rush samples back to the lab freezer
- Eliminates the need to freeze and grind most tissue samples
- Perfect for tissue collection "in the field"
- Flexible tissue storage—RNA is stable for 1 day at 37°C, 1 week at 25°C, 1 month at 4°C or long term at -20°C
- Compatible with many RNA isolation procedures including most of Ambion's RNA isolation kits
What Is RNAlater?
RNAlater is an aqueous tissue storage reagent that stabilizes and protects cellular RNA in intact, unfrozen tissue samples. RNAlater eliminates 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 for storage without jeopardizing the quality or quantity of RNA obtained after subsequent RNA isolation. RNAlater can be added to cell pellets and even cells in medium. The samples can then be stored frozen or unfrozen.
How Do You Use RNAlater?
The dissected tissue (less than 0.5 cm in any one dimension) is simply submerged in approximately 5 volumes of RNAlater (e.g., a 0.5 g sample requires about 2.5 ml of RNAlater) at room temperature. The solution permeates the cells, stabilizing the RNA. The sample can then be stored long term 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 and treated as though it had just been harvested. Most tissues can be transferred directly to a lysis buffer and homogenized, however, some hard tissues such as bone may require a more rigorous method of disruption. Samples treated with RNAlater 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.
Compatible with a Variety of RNA Isolation Procedures
RNAlater is compatible with one-step RNA isolation methods, such as TRIzol® Reagent (Invitrogen), TRI Reagent® (MRC, Ambion), RNAzol™ (Tel-Test), and RNA STAT-60™ Reagent (Tel-Test); with glass binding methods such as Qiagen's RNeasy™ or Ambion's RNAqueous®; with acid phenol extraction methods such as Ambion's ToTALLY RNA™; and with methods that use oligo(dT) selection of mRNA, such as Ambion's Poly(A)Purist™. The tissue sample is simply removed from the RNAlater and processed like fresh tissue in the RNA isolation lysis solution. Cells can be spun out of RNAlater and then added to lysis buffer, or in some cases, RNAlater can be added along with the cells directly to the lysis buffer.
Some of the Many Applications of RNAlater
RNAlater has been tested on a variety of mammalian tissues, plants, E. coli, Xenopus, fish, and Drosophila. RNAlater 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 in RNAlater to stabilize RNA during long, tedious dissections
- 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
In-house research, as well recently published independent research, indicates that the use of RNAlater for tissue storage does not affect the outcome of subsequent RNA expression analysis experiments compared to other processing methods. For an overview of this research, see Preserve Samples for RNA Expression Microarrays.
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