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Isolate It All: siRNA miRNA ,,, Total RNA Native Protein

with the mirVana PARIS Kit analyzed on an Agilent 2100 bioanalyzer.


Readily Enrich for Small RNAs
Small RNA analysis often requires the use of extremely large amounts of input RNA. Furthermore, small RNA and mRNA species are analyzed with techniques that are not compatible, for example gel purification or Northern blotting on 15% acrylamide gels for miRNA/siRNA vs. RT-PCR or microarray analysis for mRNA. With the mirVana PARIS Kit, fractions enriched in RNA species smaller than 200 nt and fractions containing only the longer RNA species can be prepared from the same experimental sample using differential binding conditions on GFF. Thus the kit allows for separate downstream analysis of protein, small RNA, and mRNA if required.

Representative data of enrichment/depletion of small RNAs obtained with the mirVana PARIS Kit are presented in Figure 3. The depleted and enriched fractions, or total RNA, were purified from the same number of HeLa cells in triplicate. Analysis of 1 g RNA on a denaturing agarose or acrylamide gel showed that 28S, 18S and 5.8S rRNA were quantitatively recovered in the fraction depleted in small RNA. In contrast, RNAs smaller than 200 nt, such as 5S RNA and tRNA, were significantly enriched in the corresponding "enriched" fraction when compared to the total RNA sample. Quantitative analysis of the same samples by Northern blot confirmed that smaller RNA species such as the microRNA miR-16 (22 nt) were also enriched ~10 fold (Figure 3B).

Figure 3. Small
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