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Invitrogen™ Quant-it™ RiboGreen Reagent and RNA Assay Kit

Catalog No. R11490
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Quantity:
1 kit
1 mL
Product Type:
Quant-iT RiboGreen RNA Kit
Quant-iT RiboGreen RNA Reagent
RediPlate 96 RiboGreen RNA Kit
Quantitation Range:
1 to 200 ng
3 to 200 ng
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Catalog No. Quantity Product Type Quantitation Range
R11490 1 kit Quant-iT RiboGreen RNA Kit 1 to 200 ng
R11491 1 mL Quant-iT RiboGreen RNA Reagent 1 to 200 ng
R32700 1 kit RediPlate 96 RiboGreen RNA Kit 3 to 200 ng
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Catalog No. R11490 Supplier Invitrogen™ Supplier No. R11490
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Description

Quant-iT RiboGreen RNA Reagent is an ultrasensitive fluorescent nucleic acid stain for quantitating RNA in solution. The Quant-iT RiboGreen RNA Assay Kit comes with reagent, dilution buffer, and rRNA standards to provide a linear detection range of 1–200 ng of RNA.

Quant-iT RiboGreen RNA Assay Kit
Containing one of the most sensitive detection dyes for the quantitation of RNA, the Quant-iT RiboGreen RNA Assay Kit and the RediPlate 96 RiboGreen RNA Quantitation Kit offer linear fluorescence detection in the range of 1–200 ng of RNA. Both assay kits contain Quant-iT RiboGreen RNA reagent as well as buffers and RNA standards.

RediPlate 96 RiboGreen RNA Quantitation Kit
The RediPlate 96 RiboGreen RNA Quantitation Kit is a rapid, simple, and highly sensitive fluorescence-based RNA-quantitation assay. The microplate is pre-loaded with RiboGreen reagent, which shows bright green fluorescence upon binding to RNA or DNA. Just add buffer and samples to the microplate wells, wait 10 minutes, and read the fluorescence in any standard fluorescence-based microplate reader. The versatile RediPlate 96 design is ideal for either high-throughput RNA quantitation or the measurement of a small sample numbers.

The RediPlate 96 RiboGreen RNA Quantitation Kit also offers the following benefits:
• Automatable—a 10 minute incubation time with no separation steps makes the assay ideal for automated, high-throughput measurements
• Precise—the average coefficient of variation for replicate assays is ≤5% using a robotic liquid handling system
• Convenient—each plate includes pre-measured RNA standards and is ready for immediate use
• Easy to read—the assay can be performed using standard fluorescence-based microplate readers

Applications
 The RediPlate 96 RiboGreen RNA Quantitation Kit is ideal for measuring yields of RNA preparations for microarray experiments, reverse transcription PCR (RT-PCR), differential display PCR, northern blot analysis, S1 nuclease assays, RNase protection assays, and cDNA library preparations.

Quant-iT RiboGreen RNA Reagent
The Quant-iT RiboGreen RNA Reagent is one of the most sensitive fluorescent dyes for the quantitation of RNA in solution, with linear fluorescence detection in the range of 1-200 ng of RNA. This reagent is also included with the Quant-iT RiboGreen RNA Assay Kit and comes complete with buffer and RNA standard.

Order Info

Shipping Condition: Room temperature

Specifications

Detection Method Fluorescence
Excitation/Emission 500/525
For Use With (Equipment) Microplate Reader
No. of Reactions 200-2,000 cuvette assays using a 2 mL assay volume
Quantitation Range 1 to 200 ng
Product Line Quant-iT, RIBOGREEN
Product Type Quant-iT RiboGreen RNA Kit
Quantity 1 kit
Shipping Condition Room Temperature

Frequently Asked Questions (FAQs)

Why am I getting negative fluorescence values with my Qubit Assays?

Negative fluorescence is a physical impossibility. It is an artifact from software autocorrecting for background signal. This means your reader is picking up and subtracting out background light at the cost of your data. Make sure to do a buffer-only control and assess the type of signal. You may need to switch to a different plate.

What are the linear ranges of the Quant-iT RNA kits?

The linear ranges are:

- Quant-iT RiboGreen RNA Assay: 1 ng/mL to 1 µg/mL
- Quant-iT RNA BR Assay: 20-1,000 ng
- Quant-iT RNA HS Assay: 5-100 ng

What is the difference between the Quant-iT RNA assay and the Quant-iT RiboGreen RNA Assay?

There are several differences:

- The Quant-iT RNA Assay uses a far-red dye (excitation 644 nm/emission 673 nm); the Quant-iT RiboGreen RNA assay uses a green fluorescent dye (excitation 500 nm/emission 525 nm).
- The Quant-iT RNA assay is more specific for RNA.
- Quant-iT RNA Assays are less sensitive than the Quant-iT RiboGreen RNA Assay.

Can I make my own assay for the Qubit Fluorometer?

Yes, you can, for Qubit instruments developed after the original Qubit (1.0) Fluorometer. See MyQubit assay instructions here (http://www.thermofisher.com/us/en/home/life-science/laboratory-instruments/fluorometers/qubit/qubit-assays/myqubit.html.html).

I have a crude lysate. Will the Quant-iT and Qubit assays work?

Generally, the cleaner the sample the better. Some salts, proteins, and detergents are tolerated in the assays; see the specific assay protocol for which ones and at what concentrations.

How does the accuracy and sensitivity of the Qubit quantitation assays using the Qubit fluorometer compare to a microplate reader?

The accuracy and sensitivity of the Qubit quantitation assays are the same as that of a microplate reader. This was a requirement during product development. The detection limits for each Qubit kit can be found on the corresponding product manual, which can be found by searching our website by keyword or catalog number.
Can the Qubit kits give an indication of sample quality in nucleic acid samples?

No. The Qubit DNA and RNA kits only quantify the amount of either DNA or RNA in the sample. The Qubit fluorometer cannot take absorbance readings to provide a A260/A280 ratio or detect protein in nucleic acid samples. This can be done with the NanoDrop instrument. If your sample contains protein or other contaminants that can affect the assay, it should be further purified.

If your sample may contain both DNA and RNA, one may use either (or both) the DNA and RNA Qubit kits and compare with samples treated with either RNase or DNase to get an accurate determination of DNA or RNA, respectively.
Can I use the Quant-iT and Qubit Kits with other fluorometers?

All Quant-iT and Qubit kits are compatible with all fluorometers and microplate readers that have the appropriate light sources and filters. You won't have access to the algorithm in the Qubit fluorometer for generating the standard curve provided by the instrument, instead, you must make a few dilutions of the highest standard DNA or RNA (Standard #2) in the Qubit kits to generate a standard curve with multiple data points.
Can I use the original Quant-iT Kits with the Qubit Fluorometer?

No, we do not recommend this. Some of the dyes in the original Quant-iT kits (those NOT listed as “for use with the Qubit fluorometer”) are not compatible with the Qubit Fluorometer. In addition, the new Quant-iT kits (labeled as “for use with the Qubit Fluorometer”) have standards formulated to be compatible with the Qubit fluorometer internal algorithms for the respective assays. The Qubit Fluorometer-compatible kits are also less expensive per assay if you are processing fewer than 20 samples at a time.
My Quant-iT RiboGreen RNA Kit (Cat. No. R11490) was shipped at ambient temperature but the recommended storage is 2-8 degrees C. Will this affect the stability or performance?

The Quant-iT Ribogreen RNA Kit (Cat. No. R11490) is routinely shipped at ambient temperature. Functional and analytic testing has shown that shipping this product at ambient temperature provides the same product quality as shipping the product on gel ice without impacting long-term stability or performance. The Quant-iT RiboGreen RNA Reagent (Component A) and Ribosomal RNA standard 16S and 23S rRNA (Component C) should be stored at 2-8 degrees C as directed in the User Guide upon receipt.