Quant-iT™ RiboGreen™ RNA Assay Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
This kit enables quantitation of as little as 1 ng/mL.	Add 1.0 mL of the aqueous working solution of the Quant-iT™ Ribo Green® reagent and incubate for 2 to 5 minutes at room temperature, protected from light.

Upstream tips
This kit enables quantitation of as little as 1 ng/mL.
Protocol tips
Add 1.0 mL of the aqueous working solution of the Quant-iT™ Ribo Green® reagent and incubate for 2 to 5 minutes at room temperature, protected from light.

Publication protocol

Animals were sacrificed and whole blood collected from the jugular vein using Microvette CB300 capillary serum collection tubes as directed by the manufacturer (Sarstedt Ltd, Leicester, UK). RNA was extracted from 50 µl of serum using TRIzol LS reagent (Life Technologies, Paisley, UK) as according to manufacturer’s instructions with minor modifications. The Caenorhabditis elegans miRNA cel-miR-39 (5′-UCACCGGGUGUAAAUCAGCUUG) was used as a synthetic spike-in control RNA oligonucleotide (IDT, Leuven, Belgium) as it has no mammalian homologue [28], [29]. After addition of TRIzol LS reagent, 3 µl of 5 nM synthetic miRNA oligonucleotide [29] was added and the samples vortexed vigorously to ensure complete mixing [2]. 20 µg of RNase-free glycogen (Roche, Burgess Hill, UK) was used as carrier to improve extraction efficiency. RNA was re-suspended in 30 µl of nuclease-free water. Where appropriate, serum total RNA was quantified using the Quant-iT RiboGreen RNA Assay Kit (Life Technologies).

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Quant-iT™ RiboGreen™ RNA Assay Kit below.

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