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

3.2.3.1. QuanTi™ RiboGreen RNA assay
Prepare serial dilution of rRNA standards (12.5 ng/mL to 200 ng/mL).

Make 70 μL aliquots of each standard and stock at -80°C for future uses.

Make 5 μL aliquots of fluorescent dye (Component A in RiboGreen kit) and stock them at -80 °C (see Note 8).

Take one set of standards and one Fluorescent Dye aliquot from freezer and thaw them at room temperature (RT) in the dark (important for the Dye) (see Note 9).

Prepare RNA sample dilutions at 1/30 in 1× TE buffer: Mix 1 μL of RNA/sample and 29 μL of 1× TE buffer for each sample.

Prepare enough working solution (WS) for all the experiment at a ratio of 1:200 dilution of Fluorescent Dye: 1× TE into a 15 mL tube (in the darkness).

Plate 15 μL of the standards (multichannel micropipette is recommended for reproducibility) in triplicate, and 15 μL of diluted samples in duplicate.

Add 15 μL of the WS into each well (standard and samples) and incubate the plate for 15 min at RT in the dark with lid.

Read the plate at 480-520 nm in a spectrophotometer (see Note 10).

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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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