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GelRed® Prestain Plus 6X DNA Loading Dye

GelRed® Prestain Plus 6X DNA Loading Dye includes our sensitive, non-toxic, non-mutagenic GelRed® dye. The 6X prestain loading dye is added to samples in place of gel loading buffer, and eliminates the need to add fluorescent DNA dye to the gel before or after electrophoresis. This is an improved version of our original 6X GelRed® prestain loading buffer, formulated to minimize DNA migration shift.

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Product Description

Features:

  • 6X loading dye includes GelRed® DNA stain
  • Combine gel loading and DNA staining in one step
  • Improved formulation to minimize DNA migration issues
  • Two blue tracking dyes that run at ~1.5 kb and ~200 bp in 1% agarose
  • Highly sensitive, non-mutagenic DNA detection

GelRed® is a sensitive, stable and environmentally safe fluorescent nucleic acid dye designed to replace the highly toxic ethidium bromide (EtBr). GelRed® Prestain Plus 6X DNA Loading Dye contains density agents, tracking dyes, and GelRed® dye. The 6X prestain loading dye is added to samples in place of gel loading buffer, and eliminates the need to add fluorescent DNA dye to the agarose gel during casting or after electrophoresis. The loading dye contains two blue electrophoresis tracking dyes that run at approximately 1.5 kb and 200 bp in a 1% agarose gel.

This product is an improved version of our original 6X GelRed® Prestain Loading Buffer (catalog number 41009) with brighter signal and more consistent DNA migration. When DNA is bound to GelRed® before electrophoresis, the ratio of dye to DNA can cause variable shifts in DNA migration, making it difficult to compare DNA fragment sizes between samples. GelRed® Prestain Plus 6X DNA Loading Dye is formulated to minimize this DNA migration shift, for greater consistency. GelRed® prestaining is simple and can avoid migration issues seen with GelRed® precast gels.

GelRed® and EtBr have virtually the same spectra, so you can directly replace EtBr with GelRed® without changing your existing imaging system. In addition, GelRed® is far more sensitive than EtBr, which cannot be used in DNA loading buffer to prestain DNA. GelRed® is compatible with downstream applications such as sequencing and cloning. It is efficiently removed from DNA by gel extraction kits or by phenol/chloroform extraction and ethanol precipitation.

GelRed® Safety
GelRed® was subjected to a series of tests at Biotium and by three independent testing services to assess the dye’s safety for routine handling and disposal. Test results confirm that the dye is impenetrable to both latex gloves and cell membranes. The dye is noncytotoxic, nonmutagenic, and classified as non-hazardous for disposal under CCR Title 22 Hazardous Waste Characterization. See the GelRed® and GelGreen® Safety Report.

Agarose Gel Staining Methods Comparison

Product / MethodProcedureAdvantagesDisadvantagesRecommended for
DNA prestaining with GelRed® Prestain Plus 6X DNA Loading DyeGelRed® loading buffer is added directly to the DNA sample before loading• Fast & simple: one-step sample loading & DNA staining

• Less concentrated dye for safer handling

• Can re-run a gel to use empty lanes

• Not recommended for PAGE, DGGE, EMSA, or PFGE gels

• Dye may cause band migration issues when loading larger amounts of DNA (more than ~100 ng/band), or for some restriction digests

• Routine agarose gels

• Recommended loading 50-200 ng ladder or 2-5 uL PCR product ( ~100 ng/band or less)
Precast gel staining with GelRed® 10,000X in waterGelRed® is mixed with molten agarose before gel castingFamiliar protocol, rapid results
Precast gel staining with GelRed® Agarose LE
Agarose is supplied pre-coated with GelRed®, just dissolve, heat, and pourSafer & more convenient, no need to handle concentrated dye
Post-electrophoresis gel staining with GelRed® 10,000X in water
- or -
GelRed® 3X in water
No fluorescent dye is added to the gel, it is stained in 3X GelRed® solution after electrophoresis• Most accurate sizing/sharpest bands

• Staining solution can be re-used

• Enhance sensitivity by adding NaCl
Extra staining step (up to 30 minutes) after electrophoresis (some customers report good results after only 5 minutes if dye is not reused)• Highly accurate band sizing

• If more than ~100 ng DNA per band must be loaded

• Analyzing restriction digests

References

Download a list of selected References for GelRed® and GelGreen®.

 

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Support & Faq

GelRed® and GelGreen® Nucleic Acid Gel Stains

The main difference between GelRed® and GelGreen® is their fluorescence excitation and emission wavelengths. GelRed® has red fluorescence, similar to ethidium bromide. GelGreen® has green fluorescence, similar to SYBR® Green or SYBR® Safe. Both dyes are compatible with standard UV transilluminators. GelGreen® is also compatible with blue light transilluminators, which allow users to avoid exposing themselves and their DNA samples to ultraviolet radiation.

GelRed® and GelGreen® have higher sensitivity for double stranded nucleic acids compared to single stranded nucleic acids, but GelRed® is more sensitive for staining single stranded nucleic acids than GelGreen®. GelRed® is about twice as sensitive for double stranded nucleic acids compared to single-stranded nucleic acids, and about five times more sensitive than GelGreen® for staining single stranded nucleic acids.

For more information about these products, please visit our DNA stains technology page.

View the GelRed® Product Line

View the GelGreen® Product Line

Category: GelRed® and GelGreen® Nucleic Acid Gel Stains

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The water formulation is a newer and improved product compared to the stock in DMSO. We recommend using dyes in water to avoid the potential hazards of handling DMSO, which can be absorbed through the skin. We continue to offer dyes in DMSO because some users do not wish to alter their established laboratory protocols. Based on internal testing, both formulations perform similarly.

Category: GelRed® and GelGreen® Nucleic Acid Gel Stains

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GelRed® is compatible with a standard UV transilluminator (302 or 312 nm).
GelGreen® has sufficient absorption between 250-300 nm and a strong absorption peak at around 500 nm. GelGreen® is compatible with a 254 nm UV transilluminator or a gel reader with visible light excitation such as a Dark Reader or a 488 nm laser gel scanner.

Category: GelRed® and GelGreen® Nucleic Acid Gel Stains

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GelRed® and GelGreen® troubleshooting

Many customers use GelRed® or GelGreen® precast gels for convenience. However, because GelRed® and GelGreen® are high affinity dyes designed to be larger dyes to improve their safety, they can affect the migration of DNA in precast gels. Some samples, such as restriction digested DNA may migrate abnormally in GelRed® or GelGreen® precast gels.

Tip #1: Load less DNA

Smearing and smiling in GelRed® or GelGreen® precast gels most often caused by overloading of DNA. If you see band migration shifts or smearing and smiling, try reducing the amount of DNA loaded. The recommended loading amount for ladders and samples of known concentration is 50-200 ng/lane. For samples of unknown concentration, try loading one half or one third of the usual amount of DNA. This usually solves band migration problems.

Tip #2: Try the post-staining protocol

To avoid any interference the dye may have on DNA migration, we recommend using the post-staining protocol. If your application requires loading more than the recommended amount of DNA, use the post-staining protocol. While we recommend post-staining gels for 30 minutes, you may be able see bands in as little as five minutes, depending on how much DNA is present. Post-staining solutions can be reused. See the GelRed® Product Information Sheet or GelGreen® Product Information Sheet for detailed protocols.

Other tips to improve agarose gel resolution:

  • If you see DNA migration issues or smearing after post-staining with GelRed® or GelGreen®, then the problem is not caused by the nucleic acid dye. Avoid overfilling gel wells to prevent smearing of DNA down the surface of the gel.
  • Pour a lower percentage agarose gel. Higher molecular weight DNA separates better with a lower percentage gel.
  • Change the running buffer. TBE buffer has a higher buffering capacity than TAE buffer.

Category: GelRed® and GelGreen® troubleshooting, GelRed® and GelGreen® Nucleic Acid Gel Stains

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There are a few possibilities:

  1. The dye may have precipitated out of solution.
    • Heat the GelRed® or GelGreen® solution to 45-50°C for two minutes and vortex to dissolve.
    • Store dye at room temperature to avoid precipitation.
  2. If you are seeing high background staining of the gel, the agarose that you are using may be of low quality. Contaminants in the agarose may bind to the dye, resulting in increased background.

Category: GelRed® and GelGreen® troubleshooting, GelRed® and GelGreen® Nucleic Acid Gel Stains

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Product shelf life, solubility, shipping and storage, and stability

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:
• Succinimidyl esters (SE, also known as NHS esters), such as our amine-reactive dyes
• Acetoxymethyl esters (AM esters) such as our membrane-permeable ion indicator dyes
• Diacetate-modified dyes, like ViaFluor™ 405, CFDA, and CFDA-SE cell viability/cell proliferation dyes

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 are generally susceptible to hydrolysis, which can result in lower labeling efficiency. Heavily sulfonated dyes, such as the Alexa Fluor® dyes, DyLight® dyes and IRDyes® are particularly hygroscopic, worsening the hydrolysis problem. For example, the percent of active Alexa Fluor® 488 succinimidyl ester (SE) could be well below 50% by the time of application (according to the manufacturer’s product datasheet). In a number of Alexa Fluor® SE reactive dyes, the SE group is derived from an aromatic carboxylic acid, while in all of Biotium’s CF™ dyes the SE group 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.

Category: Mix-n-Stain™ Antibody Labeling Kits, DNA Quantitation Kits, NucView® Caspase 3 Enzyme Substrates, Luciferase Assays, CF® Dyes, Product shelf life, solubility, shipping and storage, and stability

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Many of our solid compounds are packaged by lyophilization, in which case they usually do not appear as fluffy powders, but form a film or coating on the sides of the vial. Simply add the appropriate volume of the recommended solvent to the vial to make the desired concentration stock solution, and swirl or gently vortex to mix. Make sure the solvent comes in contact with the inside walls of the vial to fully recover the product.

Category: Product shelf life, solubility, shipping and storage, and stability

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Most of our products are stable at room temperature for many days, but we recommend storage at 4°C or -20°C to prolong shelf life. In the case of many of our aqueous dye solutions, the compounds are very stable at room temperature, but we recommend cold storage to prevent the growth of mold or other microbes over time. Therefore, to save on shipping costs, products with recommended storage at 4°C or -20°C may ship at ambient temperature or with an ice pack. These products may thaw without affecting product performance. When you receive the product, place it under the recommended storage conditions.

Category: Mix-n-Stain™ Antibody Labeling Kits, DNA Quantitation Kits, NucView® Caspase 3 Enzyme Substrates, Luciferase Assays, CF® Dyes, Product shelf life, solubility, shipping and storage, and stability

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Shipping/Shelf-life

Shipping & Handling Disclaimer: Shipping and handling methods are calculated based on item temperature storage recommendation. Large, multi-item orders may be assessed additional fees. Shipping to Puerto Rico may be assessed additional fees. A Biotium customer service representative will contact you if additional fees are required. For expedited shipping, please request in the Order Notes section of the checkout page. Please note that products with recommended storage at 4°C or -20°C may ship at ambient temperature. This will not affect product performance. When you receive the product, place it under the recommended storage conditions.