mirVana™ miRNA Isolation Kit, with phenol

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	For RNA to dissolve better and to yield 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 a higher 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. This will facilitate the removal of tightly bound proteins typically associated with RNA.

Protocol tips

For RNA to dissolve better and to yield 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 a higher 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. This will facilitate the removal of tightly bound proteins typically associated with RNA.

Publication protocol

Total RNA was extracted using the mirVanaTM kit (Life Technologies, ThermoFisher Scientific). Tissue disruption was accomplished with a TissueLyser II instrument (Qiagen, Carlsbad, CA). Sample concentration was determined using the NanoDrop® ND-1000, and RNA integrity was analyzed using the Agilent 2100 Bioanalyzer Lab-On-A-Chip Agilent 6000 Series II chip. Sample labeling and hybridization was performed per the manufacturer’s protocol. Samples were hybridized to the SurePrint G3 Mouse GE 8x60K Microarray and scanned using the Agilent G2505C Microarray Scanner. Raw data were quality-assessed, filtered for outliers and normalized to remove non-biological variation

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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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