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How to Store RNA
RNA may be stored in a number of ways. For short-term storage, RNase-free H2O ( with 0.1 mM EDTA) or TE buffer (10 mM Tris, 1mM EDTA) may be used. RNA is generally stable at -80° C for up to a year without degradation. Magnesium and other metals catalyze non-specific cleavages in RNA, and so should be chelated by the addition of EDTA if RNA is to be stored and retrieved intact. It is important to use an EDTA solution known to be RNase-free for this purpose (older EDTA solutions may have microbial growth which could contaminate the RNA sample with nucleases). It has been suggested that RNA solubilized in formamide may be stored at -20°C without degradation for at least one year (Chomczynski, 1992).
For long term storage, RNA samples may also be stored at -20°C as ethanol precipitates. Accessing these samples on a routine basis can be a nuisance, however, since the precipitates must be pelleted and dissolved in an aqueous buffer before pipetting, if accurate quantitation is important. An alternative is to pipet directly out of an ethanol precipitate that has been vortexed to create an even suspension. We have found, however, that while this method is suitable for qualitative work, it is too imprecise for use in quantitative experiments. RNA does not disperse uniformly in ethanol, probably because it forms aggregates; non-uniform suspension, in turn, leads to inconsistency in the amount of RNA removed when equal volumes are pipetted. |
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