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

6% SDS-PAGE and stained with Coomassie blue.


To further examine the compatibility of the mirVana PARIS Kit with tissues, the enrichment procedure was performed with mouse brain or kidney that were stored for several months in RNAlater, Ambion's tissue collection/stabilization solution. Previous studies showed that both high quality protein and RNA are recovered from RNAlater-treated samples [2, 6]. Analysis by denaturing acrylamide gel and Northern blot of the RNA samples purified with the mirVana PARIS Kit confirm that the procedure efficiently fractionates long and short RNA species (Figure 6). mRNAs and 5.8S rRNA were detected only in the "depleted" fraction, and the miRNAs let-7 or miR-124 were seen only in the "enriched" fraction.

Figure 6. Small RNA Enrichment from Mouse Tissues. Total RNA or fractions depleted or enriched in small RNA were isolated from RNAlater-treated mouse brain or kidney (~50 mg) with the mirVana PARIS Kit (A) 1 g of each fraction was resolved on a 15% denaturing acrylamide gel and stained with ethidium bromide. (B) mRNA, rRNA and miRNA expression was analyzed as described in Figure 4. This figure demonstrates the broad range of RNAs that can be obtained with this procedure, from miRNAs (~22 nt) to much larger mRNA species (in the kilobase range).


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In summary, these results show that high quality total RNA and protein are efficiently recovered from cultured cells and several differ
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