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Stabilizing RNA in Fresh and Frozen Tissues for RNA Isolation

RNAlater Tissue Collection: RNA Stabilization Solution for Freshly Dissected Tissue and Cells.


Problem:
Successful RNA isolation requires rapid processing or appropriate storage of the tissue or cell samples. Tissue samples must be quickly dissected on ice, followed by immediate disruption in lysis buffer. Alternatively, they can be quickly frozen using liquid nitrogen or dry ice. Any delay in processing can result in decreased RNA yield and integrity.

Solution:
RNAlater is an aqueous, non-toxic tissue and cell storage reagent that stabilizes cellular RNA within intact, unfrozen tissue and cell samples. At the same time, it "freezes" the RNA expression pattern so that analysis of the RNA will reflect actual expression levels of message at the time of sample collection. Tissue pieces submerged in RNAlater can be stored at room temperature for 1 week, 4C for a month, or -20C to -80C indefinitely. RNAlater can also be added to cell pellets and even cells in media. Samples can be shipped across continents at ambient temperature or on wet ice. RNAlater is compatible with all RNA isolation methods examined. For an example of RNAlater's use with bacteria cells, see the article Preserving Campylobacter jejuni RNA for Microarray Analysis, at right.


RNAlater-ICE Frozen Tissue Transition Solution for Frozen Tissue Specimens and Cells.

Problem:
Historically, frozen tissue samples had to remain frozen on dry ice or in liquid nitrogen prior to and during disruption in order to preserve RNA integrity. They were then ground into a powder with a chilled mortar and pestle before disruption in lysis buffer. This procedure is very laborious, especially when multiple samples must be processed, and there is risk of the tissue/powder thawing, resulting in RNA degradation.

Solution:
RNAlater-ICE addresses the issues of working with frozen tissue samples. Frozen tissue samples are simply submerged in 10 volumes of RNAlater-ICE and stored overnight at -20C. The solution remains liquid at these temperatures and the frozen tissue "thaws" in the RNAlater-ICE. RNA integrity is protected during this transition. Once treated, tissues can be used directly in standard homogenization and isolation protocols as though fresh. Alternatively, they can be stored at 20C. Treated tissues can even be brought to room temperature from 4C for a limited time for further dissection. Thus the same frozen tissue sample can be used multiple times for different experiments without compromising RNA integrity. Figure 1 shows the quality of RNA isolated from three different frozen mouse tissues that were immediately homogenized, ground with mortar and pestle, or thawed in RNAlater-ICE overnight at -20C. Both the stained gel and the resulting Northern blot demonstrate that RNA isolated from tissues treated with RNAlater-ICE maintains a high degree of integrity.

Figure 1. Quality of RNA From Samples Treated With RNAlater-ICE. Total RNA was isolated from various frozen mouse tissue samples that were either processed directly from a frozen state or thawed in RNAlater-ICE at -20C overnight. (A) shows the ethidium bromide stained RNA in a denaturing agarose gel. (B) shows the results of Northern blot analysis of the same gel hybridized to radiolabeled probes for -actin, GAPDH, and cyclophilin (Note that the specific activity of the cyclophilin probe was lower than that of the other probes). For each tissue, the integrity of both rRNA and mRNAs for the RNAlater-ICE treated samples is comparable to that of RNA prepared directly from frozen tissue samples.


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Ordering Information
Cat# Product Name Size 7020 RNAlater 100 ml 7021 RNAlater 500 ml 7022 RNAlater 50 x 1.5 ml 7023 RNAlater 20 x 5 ml 7024 RNAlater 250 ml 7030 RNAlater-ICE 25 ml 7031 RNAlater-ICE 10 x 25 ml
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