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not compatible with those appropriate for mRNA analysis. For example, small RNAs cannot be analyzed by RT-PCR or standard microarray techniques, but instead are analyzed typically by Northern blot using 15% acrylamide gels or specialized solution hybridization techniques. With the mirVana miRNA Isolation Kit, fractions enriched in RNA species smaller than 200 nt and fractions comprised of the longer RNA species can be prepared from the same sample by using differential binding conditions on the silica filters provided with the kit. This facilitates downstream analysis of both small RNA and mRNA.

A representative small RNA enrichment/depletion experiment is presented in Figure 3. The small RNA-depleted and -enriched fractions were purified from A. thaliana and compared with total RNA isolated from the same amount of tissue. Analyses on denaturing agarose and acrylamide gels demonstrate that small RNAs were quantitatively recovered in the fraction enriched for small RNAs. In contrast, longer RNA species such as the actin-2 mRNA were detected only in the fraction depleted of small RNAs. Further analysis by Northern blot showed that the same amount of microRNAs miR-163 and miR-167 were recovered in the small RNA-enriched fraction and in the total RNA sample. Since only about 200 ng of the small RNA fraction was loaded on the gel, this corresponded to a 10-fold enrichment relative to the amount of total RNA analyzed (2 g).

Figure 3. Small RNA Enrichment from A.thaliana. RNA was isolated from the same amount of tissue
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