- Immediate RNase inactivation
- Freedom from liquid nitrogen
and freezers
- Eliminates freezing and
grinding
- Perfect for field collection
- Flexible tissue storage
- Compatible with most
RNA isolation procedures including all of Ambion's RNA isolation
kits
RNAlater is an aqueous, non-toxic tissue
storage reagent that rapidly permeates tissues to stabilize and
protect cellular RNA. 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. Revolutionary reagent eliminates
the need to immediately freeze samples to preserve RNA Make Time Stand Still with RNAlater Once saturated in RNAlater, tissues
can be removed, dissected or weighed at room temperature, then
returned to RNAlater for continued storage. RNAlater-treated
tissues archived at -80°C can be subjected to 10 freeze/thaw
cycles without affecting the quality of subsequently isolated
RNA (Figure 1). Samples stored in RNAlater can be shipped
on wet ice or even at room temperature if shipped overnight.
Figure 2 shows ribonuclease protection assay data using RNA from
tissue stored in RNAlater.
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Figure 1. RNA
Stability of RNAlater Treated Tissues After Multiple
Freeze/Thaw Cycles. Mouse
heart and liver were dissected and placed in RNAlater.
The tissues were frozen on dry ice and thawed at room temperature
for the indicated number of cycles. RNA was isolated using
RNAWIZ™, electrophoresed on a 1% formaldehyde/agarose gel,
ethidium bromide stained and photographed. |
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Figure 2. Ribonuclease
Protection Assay Using Mouse Tissues Stored in RNAlater. Tissues
were stored in RNAlater for 1 or 4 weeks at 4°C.
RNA was isolated from each tissue and analyzed using the
RPA III™ Kit. |
How do I use RNAlater?
Cut tissue to be less than 0.5 cm in at least
one dimension and simply submerge it in 5 volumes of RNAlater (e.g. a 0.5 g sample requires about 2.5 ml of RNAlater).
Small organs such as rat kidney, liver and spleen can be stored
whole in RNAlater. Resuspend pelleted cells in a small
amount of PBS before adding 5-10 volumes of RNAlater.
Samples can be stored at 4°C for one month, at 25°C
for one week or at -20°C indefinitely. Archive RNAlater-treated
tissues at -20°C.
For RNA isolation, simply remove the tissue
from RNAlater 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 compatible with my system?
RNAlater has been tested on a variety
of mammalian tissues, plants, E. coli, Xenopus,
fish and Drosophila. It has been successfully used with
all of Ambion's RNA isolation kits (Figure 3) including RNAqueous™,
ToTALLY™ RNA, RNA WIZ™ and Poly(A)Pure™, as well as TRI® Reagent
(MRC). Other commercial RNA isolation kits and standard RNA isolation
protocols are also compatible with RNAlater.
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Figure 3. Compatibility
of Various RNA Isolation Methods with RNAlater. Freshly
dissected whole mouse liver and heart were placed in RNAlater and
stored at 4°C for three days. RNA was isolated from
equal mass amouts of each tissue using Ambion's ToTALLY™ RNA,
RNAqueous™ or Poly(A)Pure™ Kits. |
Does RNAlater preserve the histology
of tissues?
Recent research by Drs. Scott Florell and Sancy
Leachman at the Huntsman Cancer Institute [Florell et al. (2001) Mod
Pathol 14:116-128] demonstrate that RNAlater does
preserve the histology of tissues. A blinded study was performed,
in which 2 pathologists compared human tissue sections that were
either immediately processed for histology (fixed in formalin
and embedded in paraffin, or frozen sectioned) to samples stored
in RNAlater, rinsed, and then processed for histology.
Their results indicate that morphological detail and staining
characteristics were identical for the 2 groups of samples. An
example of their findings is presented in Figure 4. In addition
to excellent staining and preservation of morphological detail,
many immunohistological stains performed equally well in the
RNAlater preserved samples, suggesting preservation in
RNAlater caused no damage to cellular epitopes.
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| Figure 4. Histology
of RNAlater Preserved Tissue. RNAlater preserved
samples are suitable for histology and provide excellent
morphological detail. Sections made from RNAlater preserved
material were indistinguishable from slides made from untreated
samples when examined for standard histological criteria.
Panel A. H&E stained, frozen section of human skin preserved
for one week in RNAlater prior to processing. Panel
B. Stained as above but formalin-fixed, paraffin-embedded
section of human skin preserved for one week in RNAlater prior
to processing. |
How practical is RNAlater?
This revolutionary and inexpensive product
preserves the integrity of your precious RNA and saves you the
time and trouble of grinding frozen tissue in liquid nitrogen.
RNAlater is economical and can be stored at room temperature.
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