Invitrogen™ Quant-iT™ PicoGreen™ dsDNA Assay Kits and dsDNA Reagents

Product Code.	Quantity	Product Type	Packaging Type	unitSize
11559196	10 x 100 μL	dsDNA Reagent	10 x 100 μL Tubes	1000µL
10545213	1 mL kit	dsDNA Assay Kit	1 x 1 mL Bottle	1mL
10398702	10 x 100 μL kit	dsDNA Assay Kit	10 x 100 μL Tubes	Each
10535213	1 mL	dsDNA Reagent	1 x 1 mL Bottle	1mL

Product Code. 11559196 Supplier Invitrogen™ Supplier No. P11495

Description

Selectively detect dsDNA from a wide variety of sources, and even in presence of ssDNA, RNA, and free nucleotides, with the Quant-iT PicoGreen dsDNA Assay Kit and PicoGreen dsDNA Reagent.

Achieve precise and accurate dsDNA concentration measurements over a broad dynamic range with the Quant-iT Picogreen dsDNA Assay kits and dsDNA reagents. Picogreen assays are compatible with most microplate readers and fluorometers.

Rapidly detect as little as 0.25 pg/uL of dsDNA, even in the presence of ssDNA, RNA, and free nucleotides, using the PicoGreen dsDNA quantitation assay and reagent. The assay is linear over three orders of magnitude and has little sequence dependence, allowing you to accurately measure DNA from many sources, including genomic DNA, viral DNA, miniprep DNA, or PCR amplification products.

PicoGreen dsDNA quantitation reagents are
• Significantly more sensitive than UV absorbance readings, saving precious sample amounts
• Specific for dsDNA in the presence of RNA
• Easy to use—just add the dye working solution to the sample, incubate five minutes, and read
• Suitable for use in 96- and 384-well plate formats
• Compatible with most fluorescence-based microplate readers and fluorometers
• Picogreen aproximate fluorescence absorption/emission maxima is 502/523 nm, bound to dsDNA

Applications
The PicoGreen dsDNA quantitation reagent and kits are ideal for PCR-based assays, microarray samples, DNA damage assays, enzyme activity assays, genomic DNA quantitation, measuring dsDNA in complex mixtures, and viral DNA quantitation.

Specifications

• Detection Method: Fluorescence
• Excitation/Emission: 500/525
• For Use With (Equipment): Microplate Reader
• No. of Reactions: 200 Assays (2 mL assay volume)
• Packaging Type: 10 x 100 μL Tubes
• Quantitation Range: 50 pg to 2 μg
• Product Line: PICOGREEN, Quant-iT
• Product Type: dsDNA Reagent
• Quantity: 10 x 100 μL
• 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.

I have a Quant-iT DNA Kit and want to use it for the Qubit Fluorometer. Can I?

Yes, the manual has directions for this application. You will use the 0 ng/µL lambda dsDNA HS standard to generate Standard #1. You will prepare a dilution of the 10 ng/µL lambda dsDNA HS standard to generate Standard #2. You then prepare the samples and compare them to this 2-point standard curve. The Quant-iT dsDNA BR Kit can be used in a similar manner.

What is the useful pH range for Quant-iT DNA kits?

The buffer included in the kit should assure the proper pH range, even if your DNA is at a pH outside of this range, since at least a 10-fold excess of kit buffer over sample is used in the assay.

I'm trying to quantify some DNA labeled with a fluorophore. Will this work?

PicoGreen dye and other fluorescence-based quantification reagents are not recommended for quantifying dye-conjugated nucleic acids. The attached dye molecules can interfere with either binding and/or fluorescence output of the quantification reagents.

Does DNA length have an effect on the dsDNA assays?

Strands that are roughly in the 20-mer range or shorter show a lower level of signal. For dsDNA samples that are composed of mostly short strands, the reagent may still be used, but one should use a dsDNA standard that is of comparable length as the sample.

I want to adapt my Quant-iT PicoGreen DNA kit to a 96-well plate format. Is this possible?

Yes, we would recommend reducing the volume while keeping the dye: sample ratio constant.

What is the difference between the Quant-iT PicoGreen DNA, Quant-iT DNA, and Qubit DNA assays?

The Qubit Fluorometer contains highly optimized algorithms that calculate the concentration of the sample using either the Qubit assays or the Quant-iT DNA assays. The Quant-iT PicoGreen DNA assay may be adapted to the Qubit Fluorometer using the MyQubit firmware. The performance of all of these assays is similar.

The Quant-iT PicoGreen DNA assay is the most established assay and the most general-purpose (http://tools.thermofisher.com/content/sfs/manuals/PicoGreen-dsDNA-protocol.pdf). It requires the dilution of the standard DNA and buffer but can be adapted for use with either cuvettes, microplates, or the NanoDrop 3300.

The Quant-iT DNA assays provide a ready-to-use buffer and pre-diluted standard DNA for analyzing a large number of samples (>20 samples) using a 96-well microplate with no further adaptation.

The Qubit assays (https://www.thermofisher.com/us/en/home/industrial/spectroscopy-elemental-isotope-analysis/molecular-spectroscopy/fluorometers/qubit/qubit-assays/myqubit.html) are intended for low throughput (<20 samples), and are only used on the Qubit Fluorometer.

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.

I want to see mitochondrial DNA. Which dye is best?

Several nucleic acid stains will label mitochondrial DNA as well as nuclear DNA. One stands out above the rest: PicoGreen stain. It will label nuclear DNA as well, but the mitochondrial DNA label stands out in cells that are adherent and relatively large. See the publication: Exp Cell Res. (2005) 303(2):432-46 .

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.