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Are there any alternatives for ethidium bromide in agarose gels?

Posted December 17, 2021


Answer

These are some of the more widely used alternatives for ethidium bromide in agarose gels include:

SYBR Safe

SYBR Safe is as sensitive as Ethidium Bromide but less hazardous. Another advantage of using SYBR Safe is that it can be visualized using blue light excitation, which does not cause DNA damage. It binds to the DNA double helix and is used as an in-gel stain for DNA detection.

Methylene Blue
Methylene blue is not as sensitive as Ethidium Bromide because of its weaker interaction with DNA and RNA. Its two main advantages are that it is detectable in the visible range and it is less hazardous.

SYBR Green

SYBR Green binds to DNA through two mechanisms – DNA intercalation and external binding. It binds strongly to double-stranded DNA and with lesser intensity to single-stranded DNA and RNA.

Crystal Violet

Crystal violet binds to DNA in the same way as ethidium bromide but is less mutagenic. Another advantage of using crystal violet is that it is detectable in the visible range, which reduces the risks associated with UV exposure.

Gel Red
Gel Red is a safe, sensitive, and stable DNA stain. It is visualized using UV light like ethidium bromide. However, it is less mutagenic.

Eva Green

Eva Green has a high affinity for double-stranded DNA and is virtually non-cytotoxic and non-mutagenic. It is very stable at high temperatures and fluoresces brightly on binding to dsDNA. 

Gelite Safe

As a membrane-impermeant dye Gelite Safe has extremely low to no cytotoxicity or mutagenicity. The notable features of this DNA stain are its broad excitation and emission profiles and its exceptional sensitivity and versatility.

Additional resources

Visualization of DNA in agarose gels as migrating colored bands: applications for preparative gels and educational demonstrations

Helixyte™ Green dsDNA Quantifying Reagent *200X DMSO Solution*