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FirstChoice® Total
RNA: Human Normal Tissue
FirstChoice® Total
RNA: Mouse Normal Tissue
FirstChoice® Total
RNA: Rat Normal Tissue
As research interest in the expression patterns
of miRNA increases it is important to recognize that the method
of RNA isolation utilized is a critical factor. Most RNA isolation
protocols were developed for longer messenger RNA and are not optimized
to recover small molecules. In fact, using standard glass fiber filter
of silica matrix methodologies may result in the loss of almost all
small RNA. This also has implications for commercially available purified
RNA products. Depending on the isolation procedures that were employed,
many of these products may not contain small RNA. To address
this problem Ambion has validated our isolation procedures to efficiently
recover small RNA molecules. As such, we can now certify that the FirstChoice
Total RNA products listed above contain miRNA.
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| Detection
of miRNAs Across Different Tissues. (A) miR-16,
miR-20 and miR-22 miRNAs were detected in 1 µg of FirstChoice
Total RNA from 10 different mouse tissues. The same differential
expression of miR-16 across tissues was observed by Northern
blot analysis (2 days exposure) or by hybridization in solution
with the mir Vana™ miRNA Detection Kit (2 hr
exposure). As a control, the same RNA samples (1 µg) were also resolved
on a 1.2 % denaturing agarose gel and U1 snRNA, ß-actin and GAPDH mRNA
expression was analyzed by Northern blot using the NorthernMax®-Gly Kit. (B) miR-200b,
miR-16 and miR-22 miRNAs were detected in 1 µg of FirstChoice Total
RNA from 12 different human tissues. miR-200b and let-7 expression was also
analyzed by Northern blot (4 µg of total RNA). |
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