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Content #1
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A fluorescent dsDNA quantitation kit with a linear range of 1-3,000 pg DNA, an unprecedented level of sensitivity in dsDNA detection. Designed for use in a fluorescence microplate reader.
The AccuBlue® NextGen dsDNA Quantitation assay is the most sensitive and accurate DNA quantitation method for ultra low amounts of DNA, with a linear range from ~1 – 3,000 pg (depending on instrument capability). This assay is designed for use with fluorescence microplate readers. The AccuBlue® NextGen assay is also available without any DNA standards (31061), for those that wish to use their own standard.
*Note: In order to go green and save on packaging and shipping, we have changed from a 1X to a 20X buffer.
With a detection limit as low as 1 pg DNA in a 96-well plate reader assay (see Table 1), AccuBlue® NextGen outperforms rivals such as PicoGreen® in sensitivity and accuracy. Specially designed for scientists who need to quantify extremely dilute or precious DNA, AccuBlue® NextGen is ideal for use in preparing samples for applications such as Next Generation Sequencing (NGS) and digital PCR.
Because the AccuBlue® NextGen assay is so sensitive, results will vary depending on many factors, such as microplate reader (see table below), assay volume, and pipet accuracy.
| Plate reader | Assay format (assay volume) | Lowest DNA detected |
|---|---|---|
| BMG CLARIOStar | 96-well (200 uL) | 1 pg |
| BMG CLARIOStar | 384-well (50 uL) | 0.6 pg |
| Biotek Synergy H1m | 96-well (200 uL) | 5 pg |
| SpectraMax Gemini XS | 96-well (200 uL) | 5 pg |
| SpectraMax i3 | 384-well (50 uL) | 1.25 pg |
AccuBlue®, AccuClear® and AccuGreen™ dsDNA quantitation assays allow precise quantitation of purified dsDNA samples across a wide range of concentrations and a variety of fluorescence detection instruments. Unlike absorbance-based nucleic acid quantitation, fluorescent DNA binding dyes are highly sensitive and selective for double-stranded DNA and provide a more accurate DNA concentration in the presence of contaminating RNA and other common contaminants including free nucleotides, protein, detergents and salts.
Biotium offers dsDNA quantitiation kits and solutions for different instruments and sample concentration ranges. See the table below and visit the DNA & RNA Quantitation Technology Page for details on our full line of dsDNA and RNA quantitation kits.
| Kit | DNA or RNA | Detection range (in assay)* | Dye Ex/Em (nm) | Suggested instrument | Features |
|---|---|---|---|---|---|
| AccuGreen™ High Sensitivity DNA | DNA | 0.1-100 ng | 502/523 | Qubit® fluorometer | Compare to the Qubit® dsDNA HS assay from Thermo Fisher |
| AccuGreen™ Broad Range DNA | DNA | 2-1000 ng | 500/530 | Qubit® fluorometer | Compare to the Qubit® dsDNA BR assay from Thermo Fisher Non-toxic & non-mutagentic DNA quantitation dye |
| AccuBlue® NextGen DNA | DNA | 1-3000 pg** | 468/507 | Fluorescence microplate reader | Most sensitive assay available for quantitation of precious or dilute samples Optimal for sensitive applications such as NGS or digital PCR |
| AccuClear® Ultra High Sensitivity DNA | DNA | 0.03-250 ng | 468/507 | Fluorescence microplate reader | Versatile kit with high sensitivity and wide linear range |
| AccuBlue® High Sensitivity DNA | DNA | 0.2-100 ng | 485/530 | Fluorescence microplate reader | Non-toxic & non-mutagentic DNA quantitation dye |
| AccuBlue® Broad Range DNA | DNA | 2-2000 ng | 350/460 | Fluorescence microplate reader | Broad linear range with blue fluorescence |
| AccuBlue® Broad Range RNA | RNA | 5-1000 ng | 650/670 | Fluorescence microplate reader or Qubit® fluorometer | The widest linear range of available RNA quantitation kits Exceptional accuracy, sensitivity, and high RNA selectivity |
1. Analytical Biochemistry (2016) 15 Oct pp74-79 doi:10.1016/j.ab.2016.07.022
2. J Assist Reprod Genet (2018) 35, 1497-1501 doi:10.1007/s10815-018-1223-4
1. Analytical Biochemistry (2016) 15 Oct pp74-79 doi:10.1016/j.ab.2016.07.022
2. J Assist Reprod Genet (2018) 35, 1497-1501 doi:10.1007/s10815-018-1223-4
Bioscience kits
The guaranteed shelf life from date of receipt for bioscience kits is listed on the product information sheet. Some kits have an expiration date printed on the kit box label, this is the guaranteed shelf life date calculated from the day that the product shipped from our facility. Kits often are functional for significantly longer than the guaranteed shelf life. If you have an older kit in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the kit still works for your application before processing a large number of samples or precious samples.
Antibodies and other conjugates
The guaranteed shelf life from date of receipt for antibodies and conjugates is listed on the product information sheet. Antibodies and other conjugates often are functional for significantly longer than the guaranteed shelf life. If you have an older conjugate in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples.
For lyophilized antibodies, we recommend reconstituting the antibody with glycerol and antimicrobial preservative like sodium azide for the longest shelf life (note that sodium azide is not compatible with HRP-conjugates).
Chemicals, dyes, and gel stains
Biotium guarantees the stability of chemicals, dyes, and gel stains for at least a year from the date you receive the product. However, the majority of these products are highly stable for many years, as long as they are stored as recommended. Storage conditions can be found on the product information sheet or product safety and data sheet, material safety data sheet, and on the product label. Fluorescent compounds should be protected from light for long term storage.
If you have a Biotium compound that has been in storage for longer than one year that you wish to use, we recommend performing a small scale positive control experiment to confirm that the compound still works for your application before processing a large number of samples or precious samples.
Expiration date based on date of manufacture (DOM)
If your institution requires you to document expiration date based on date of manufacture for reagents, please contact techsupport@biotium.com for assistance.
Chemical products with special stability considerations:
Esters
Ester compounds include the following:
Ester dyes are stable in solid form as long as they are protected from light and moisture. Esters are not stable in aqueous solution. Concentrated stock solutions should be prepared in anhydrous DMSO (see Biotium catalog no. 90082). Stock solutions in anhydrous DMSO can be stored desiccated at -20°C for one month or longer. Esters should be diluted in aqueous solution immediately before use. Succinimidyl esters (SE) should be dissolved in a solution that is free of amine-containing compounds like Tris, glycine, or protein, which will react with the SE functional group. AM esters and diacetate compounds should be dissolved in a solution that is free of serum, because serum could contain esterases that would hydrolyze the compound.
A note on CF® Dye succinimidyl ester stability
Succinimidyl esters (SE) are generally susceptible to hydrolysis, which can result in lower labeling efficiency. Many commercially available fluorescent dyes used for life science research are heavily sulfonated dyes which makes them particularly hygroscopic, worsening the hydrolysis problem. In addition, for several commercially available SE reactive dyes, the SE group is derived from an aromatic carboxylic acid, while the SE group in all of Biotium’s CF® Dyes is prepared from an aliphatic carboxylic acid. This structural difference reduces the susceptibility of CF® Dye SE reactive groups to hydrolysis, resulting in relatively stable reactive dyes with consistently higher labeling efficiency compared to other SE derivatives of other fluorescent dyes.
Maleimides, MTS and thiosulfate dyes
Like the succinimidyl ester dyes, these dyes are also susceptible to hydrolysis, although generally to a much lower degree. Thus, for long term storage, anhydrous DMSO is recommended for making stock solutions.
Other reactive dyes
Amines, aminooxy (also known as oxylamine), hydrazide, azide, alkyne, BCN, and tyramide reactive dyes, as well as dye free acids, are generally stable in aqueous solution when stored at -20°C for 6-12 months or longer, as long as no compounds are present that may react with the dye’s functional group. See the product information sheets for specific reactive dyes more information.
Coelenterazines and D-luciferin
Coelenterazines are stable in solid form when stored as recommended; they are not stable in aqueous solution. Concentrated coelenterazine stock solutions (typically 1-100 mg/mL) should be prepared in ethanol or methanol; do not use DMSO or DMF to dissolve coelenterazines, because these solvents will oxidize the compounds. Ethanol or methanol stocks of coelenterazine can be stored at -20°C or below for six months or longer; alcohol stocks may evaporate during storage, so use tightly sealing screw cap vials and wrap the vials with Parafilm for long term storage. Propylene glycol also can be used as a solvent to minimize evaporation. If the solvent evaporates, the coelenterazine will still be present in the vial, so note the volume in the vial prior to storage so that you can adjust the solvent volume to correct for evaporation if needed. Prepare working solutions in aqueous buffers immediately before use. Coelenterazines are stable for up to five hours in aqueous solution.
Aquaphile™ coelenterazines are water soluble formulations of coelenterazines. They are stable in solid form when stored as recommended. Aquaphile™ coelenterazines should be dissolved in aqueous solution immediately before use. They are stable for up to five hours in aqueous solution.
Note that coelenterazines are predominantly yellow solids, but may contain dark red or brown flecks. This does not affect product stability or performance. If your coelenterazine is uniformly brown, then it is oxidized and needs to be replaced.
D-luciferin is stable in solid form and as a concentrated stock solution when stored as recommended; it is not stable at dilute working concentrations in aqueous solution. Prepare concentrated D-luciferin stock solutions (typically 1-100 mg/mL) in water, and store in aliquots at -20°C or below for six months or longer. Prepare working solutions immediately before use.
Note that the optimal excitation and emission maxima of AccuBlue® NextGen are 468/507 nm which is different than the other AccuBlue® dyes. The gain settings on your fluorescence plate reader may need to be adjusted for optimal signal to background.
Any of our DNA and RNA quantitation kits can be used with the CLARIOstar reader, and we routinely test them on that model. For example, the CLARIOstar gives excellent results with the AccuClear® dsDNA Quantitation Kit, which has the broadest dynamic range, as well as the.AccuBlue® NextGen dsDNA Quantitation Kit, which has the highest sensitivity.
You can find information about our full selection of assays on the dsDNA Quantitation Technology page.