Where can I find the expiry date of GelRedTM?
Biotium guarantees the stability of GelRed TM for at least a year from the date you receive your product. However, the majority of Biotium products are highly stable for many years, as long as they are stored as recommended. Storage conditions can be found on the GelRed TM 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 GelRed TM 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.
What is the difference between GelRed™ and GelGreen?
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.
How do I use GelRed ™?
GelRed TM can be added to agarose during gel casting at a final concentration of 1X, or it can be used for post-electrophoresis gel staining at a final concentration of 3X in water. For detailed protocol please download the GelRed™ Product Information Sheet.
How many gels can I stain with a vial of GelRed?
GelRed TM can be added to agarose during gel casting at a final concentration of 1X, or used for post-electrophoresis gel staining at a final concentration of 3X in water. A 0.5 mL vial of GelRed TM can be used to prepare 100 minigels (50 mL each) using the precast protocol, or for post-electrophoresis staining of 33 minigels in 50 mL staining solution per gel. Post-staining solution also can be re-used for staining two or more gels. Many customers use GelRed™ precast gels for convenience. However, because GelRed™ is a high affinity & larger dye designed for improved safety, it can affect the migration of DNA in precast gels. Some samples, such as restriction digested DNA may migrate abnormally in GelRed™ precast gels. The following modifications may improve band resolution in precast gels.
Why am I seeing smeared or smiling DNA band(s) or discrepant DNA migration?
Many customers use GelRed™ 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™ precast gels. The following modifications may improve band resolution in precast gels.
- Reduce the amount of DNA loaded. Smearing and smiling is often caused by overloading of DNA. 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.
- 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.
- 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.
Why do I see weak fluorescence, decreased dye performance over time, or a film of dye remaining on the gel after post-staining?
The dye may have precipitated out of solution.
- Heat GelRed™ solution to 45-50°C for two minutes and vortex to dissolve.
- Store dye at room temperature to avoid precipitation.
Does post-staining require a de-staining step?
No, but de-staining with water can be performed if background is high.
Can GelRed™ be used to stain ssDNA or RNA?
GelRed™ can be used to stain both ssDNA and RNA. GelRed™ is about 5 times more sensitive for single-stranded nucleic acids than GelGreen™. Titration assays using a fluorescence microplate reader showed that the fluorescence signal of GelRed™ bound to ssDNA and RNA is about half that of GelRed bound to dsDNA.
What instruments can be used to detect GelRed™?
GelRed™ is compatible with a standard UV transilluminator (302 or 312 nm).
What emission filters are suitable for use with GelRed™?
Use the ethidium bromide filter for GelRed™. SYBR or Gelstar filters also can be used for gel imaging with equally good results. Alternatively, a long-pass yellow filter can be used with GelRed™. Please review the emission spectrum for GelRed™ for more specific wavelengths.
Can I make GelRed gels ahead of time and store them for later use?
You can store precast GelRed™ gels for up to a week, and GelGreen™ gels for up to a month. We recommend storing gels at room temperature in the dark. Storing Gels at at 4°C is no longer recommended, because this can lead to dye precipitation and poor performance.
What is the stability of GelRed™ in molten agarose?
We do not recommend storing GelRed™ in molten agarose for more than a few days.
Can I reuse a GelRed™ precast gel after running samples?
No. We do not recommend that GelRed™ gels be reused after electrophoresis because the staining intensity can be decreased with subsequent electrophoresis.
Can I re-melt a GelRed™ gel and cast again?
Yes, but it may be necessary to add some more dye to the re-melted gel for the best signal.
How should I dispose of GelRed™?
GelRed™ and GelGreen™ passed the EPA regulated Title 22 test. Some facilities have approved the disposal of these stains directly down the drain. However, because regulations vary, please contact your safety office for local disposal guidelines. GelRed™ can be adsorbed to activated carbon (also known as activated charcoal) for disposal as chemical waste. Please review the GelRed/GelGreen safety report for more detailed information.
How safe is GelRed™, and where can I find more information about its safety?
In AMES and related tests, GelRed™ was shown to be much safer alternatives to EtBr and SYBR dyes. Nevertheless, please exercise safe laboratory practices when using GelRed™.
A comprehensive safety report is available for download here: safety report.
What loading buffers are compatible with GelRed?
A routinely used 6X loading buffer contains 15% glycerol, 7.5% Ficoll 400, 0.05% Bromophenol Blue, and 0.1% Patent Blue VF. In internal testing, 6X loading buffer containing 0.1% Orange G produced good results in precast and post-stained GelRed™ gels. SDS in loading buffer may contribute to band smearing in precast GelRed™ gels. If this occurs, we recommend using the post-staining protocol.
What is the lower detection limit of GelRed™?
GelRed™ is an ultra-sensitive dye. Some users have reported being able to detect bands containing less than 0.1 ng DNA. However, the sensitivity of the staining will depend on the instrument capability and exposure settings.
What is the binding mechanism of GelRed™?
GelRed™ most likely binds via a combination of intercalation and electrostatic interaction.
(http://link.springer.com/article/10.1007/s00249-014-0995-4).
In which direction does GelRed™ migrate during electrophoresis?
GelRed™ does not migrate through the gel as easily as ethidium bromide. It is not necessary to add additional dye to the running buffer, and the gel will be stained more homogenously than a gel stained with ethidium bromide.
Does GelRed™ need to be used in the dark?
GelRed™ is very stable and can be used in room light. However, we recommend long term storage of GelRed™ in the dark between uses. A customer has reported using GelRed™ with success after accidentally leaving it in ambient light for one month.
How much GelRed™ do I need to use?
Dilute the 10,000X stock 10,000-fold for 1X precast gels (for example, 5 uL for a 50 mL gel), or 3,333-fold for a 3X post staining solution (15 uL for a 50 mL solution).
Can GelRed™ post-staining solution be reused?
Yes. However, if the sensitivity decreases, use a fresh solution of the dyes.
Is GelRed™ compatible with alkaline gel running buffer (30mM NaOH, 1mM EDTA)?
Yes.
Why is GelRed™ available in either DMSO or water?
The GelRed™ water formulation is a newer and improved product compared to the stock in DMSO. We recommend using GelRed™ in water to avoid the potential hazards of handling DMSO, a solvent which can be absorbed through the skin. Biotium continues 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.
GelRedTM and its uses are covered by US patent numbers 796048, 7803943, and 8232050.
Materials from Biotium are sold for research use only, and are not intended for food, drug, household or cosmetic use.
© Copyright 2023 Gene Target Solutions Pty Ltd