Super GelRed^® is a sensitive, stable and environmentally safe fluorescent nucleic acid dye designed to replace the highly toxic ethidium bromide (EtBr) for staining dsDNA, ssDNA or RNA in agarose gels. Super GelRed^® and EtBr have virtually the same spectra , so you can directly replace EtBr with Super GelRed^® without changing your existing imaging system. In addition, Super GelRed^® is far more sensitive than EtBr .

1.Due to the high sensitivity of Super GelRed^® and eliminating the possibility of dye interference with DNA migaration in precast, it is recommend to reduce the amount of DNA loaded. We recommend to load 50-200 ng/lane (small lanes of 8-lanes) for known concentrations of sample. For samples with unknown concentration, Try 1/3 or 1/5 of the commonly used sample volume. Appropriately increase or decrease the sample volume according to the size of the gel hole.

2.For better results, it is recommended to replace TAE with 1×TBE buffer, because the borate-containing reagents are more conductive. The voltage during electrophoresis should not be too high. Generally, TBE should not exceed 120V. TAE should not exceed 100 V.

◆ Smeared DNA bands in precast gel?

1. Reduce the amount of DNA loaded by one-third to one-fifth. Super GelRed® is much more sensitive than EtBr. Blown out or smeared bands can be caused by overloading. This is frequently observed with DNA ladders.

2. Perform post-staining instead of pre-casting.

3. Pour a lower percentage agarose gel for better resolution of large fragments.

4. Change the running buffer. TBE buffer has a higher buffering capacity than TAE.

5. Loading buffers containing SDS may contribute to band smearing. If this occurs, use the post-staining protocol for applications requiring SDS-containing loading buffers.

◆Discrepant DNA migration in pre-cast gel？

Super GelRed^® is designed to be larger than other dyes to prevent it from entering cells, thus rendering the dye safer. The migration of DNA may be affected depending on the dye: DNA ratio.

1. Reduce the amount of DNA loaded by one-third to one-fifth.

2. Post-stain gel in 3× Super GelRed® to avoid any interference the dye may have on migration during electrophoresis.

◆Weak fluorescence, decreased dye performance over time, or film of dye remains on gel after post-staining？

The dye may have precipitated out of solution.

1. Heat Super GelRed^® solution to 45-50℃ for two minutes and vortex to redissolve.

2. Store dye at room temperature to avoid precipitation.

◆Can Super GelRed^® be used to stain ssDNA or RNA？

Super GelRed^® can be used to stain ssDNA and RNA, but it is twice as sensitive for dsDNA than for ssDNA or RNA.

◆What emission filters are suitable for use with Super GelRed^®?

Use the ethidium bromide filter for Super GelRed^®. SYBR or GelStar filters also can be used for gel imaging with equally good results. Please review the emission spectra for Super GelRed^® for specific wavelengths.

◆Can I reuse a Super GelRed^® precast gel after electrophoresis?

We do not recommend reusing Super GelRed^® precast gels as signal decreases with subsequent electrophoresis.

◆How should I dispose of Super GelRed^®?

Some facilities have approved the disposal of Super GelRed^® directly down the drain. However, because regulations vary, please contact your safety office for local disposal guidelines. Super Super GelRed^® can be adsorbed to activated carbon (also known as activated charcoal) for disposal as chemical waste.

◆ What is the lower detection limit of Super GelRed^®?

Some users have reported being able to detect bands containing less than 0.1 ng DNA. However, the limit of detection will depend on instrument capability and exposure settings.

◆What is the chemical structure of Super GelRed^®?

The chemical structure of Super GelRed^® is proprietary.

◆What is the binding mechanism of Super GelRed^®?

Super GelRed^® has been shown to bind DNA exclusively by electrostatic attraction.

◆Does Super GelRed^® migrate during electrophoresis?

Super GelRed^® does not migrate through the gel as easily as EtBr. It is not necessary to add dye to the running buffer, and the gel will be stained more homogeneously with Super GelRed^® than with EtBr.

◆Does Super GelRed^® need to be used in the dark?

Super GelRed^® is very stable. You can use the dye in room light.

◆Is there a difference between 10,000× Super GelRed^® in DMSO and water?

The Super GelRed^® stock in water is a newer and improved product compared to the stock in DMSO. We recommend using Super GelRed^® in water to avoid the potential hazards of handling DMSO, a solvent that can be absorbed through the skin. We continue to offer Super GelRed^® in DMSO because some users do not wish to alter their established laboratory protocols.