mirVana™ miRNA Isolation Kit, with phenol

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- For RNA to dissolve better and to yeild high levels, preheat elution buffer at 95C. - Addition of β-mercaptoethanol in lysis buffer can be skipped because the phenol will do the job of nuclease inactivation. - To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step.	- Do vortex for 2-plus minutes during Acid-Phenol:Chloroform extraction step. This will facilitate the removal of tightly bound proteins typically associated with RNA.

Protocol tips
- For RNA to dissolve better and to yeild high levels, preheat elution buffer at 95C. - Addition of β-mercaptoethanol in lysis buffer can be skipped because the phenol will do the job of nuclease inactivation. - To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step.
Downstream tips
- Do vortex for 2-plus minutes during Acid-Phenol:Chloroform extraction step. This will facilitate the removal of tightly bound proteins typically associated with RNA.

Publication protocol

Total RNA was purified using the mirVana™ miRNA Isolation Kit (ABI, Carlsbad, CA USA). For miRNA profiling, we utilized the 5th generation (human, mouse, rat) miRCURY LNA™ microRNA Array Kit (Exiqon Inc., Woburn, MA USA). Total RNA recovered from three independent experiments were quality checked using 1% formaldehyde-agarose gel electrophoresis and an Agilent bioanalyzer, and the microarrays were processed (i.e. samples labeled, arrays hybridized, washed and scanned) by the UCLA Clinical Microarray Core facility (http://www.pathology.ucla.edu/cmc).

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for mirVana™ miRNA Isolation Kit, with phenol below.

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