MIDORI Green Advance — Safe DNA Stain

A safe alternative to ethidium bromide
MIDORI Green Advance can stain single- and double-stranded nucleic acids with sensitivity similar to that of ethidum bromide(EtBr). Like EtBr, Midori Green Advance is added directly to molten agarose during the gel pouring step. This makes it a no-brainer as a drop-in replacement for your lab’s current protocols. Not only can you use your existing UV transilluminator and filter sets — MIDORI Green Advance works best when matched with blue LED-, or in particular, Blue/Green LED-based illuminators and gel documentation systems. These environmentally-friendly light tables are the perfect complement for any MIDORI Green stain, as you can work shield-less and goggle-less when excising bands or imaging migration. And because blue or green light does not cross-link DNA, you never need to worry about damaging DNA during cloning and photographing sessions.

Comparison of sensitivity between MIDORI Green Advance (left green) and Ethidium Bromide (right red) using a Blue/Green LED Transilluminator.

MIDORI Green Advance shows a very high sensitivity even for small DNA fragments. The dilution factor of MIDORI Green Advance can be as high as 1:25000. Hence, 4 μL to 6 μL are enough for a 100 mL agarose gel, resulting in about 17 to 25 liters of stained agarose gels per 1 mL tube of MIDORI Green Advance.

The absorbance spectrum of Ethidium Bromide, MIDORI Green Advance, and MIDORI Green Direct

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Thoroughly tested for your safety
Non-toxic. Non-mutagenic. MIDORI Green Advance Nucleic Acid Stain is our most thoroughly tested alternative to traditional ethidium bromide (EtBr) stain. Five different tests show just how safe it is. As compared with EtBr, known to be a strong mutagen; MIDORI Green Advance causes far fewer mutations in the Ames test, the accepted industry standard for identifying mutagens. High mutagenicity in the Ames test is often linked to carcinogens and teratogens. MIDORI Green Advance also scores negative in the mouse bone marrow micronucleus test and in the chromosome aberration test, which indicates that it’s considered non-carcinogenic — unlike EtBr. The other side of safety is toxicity, and all MIDORI Green stains have tested non-toxic too! So these stains do not need to be treated as hazardous waste; unlike EtBr and some other “safe DNA dyes”, and can be disposed of according to standard laboratory procedures, making lab use easy and safe for technical staff and students. And for those who prefer to be extra safe, MIDORI Green Advance also tests negative for latex/nitrile glove penetration—even after 6 hours of exposure.

Three makes a happy family
The core MIDORI Green molecule has been formulated into three different green fluorescent stains optimized for your lab’s needs. First, there’s MIDORI Green Advance, which offers an excellent signal-to-noise ratio and solid sensitivity for even short nucleic acid fragments. MIDORI Green Advance is added into the agarose gel prior to running samples (much like EtBr), and provides sensitivity of detection on par with EtBr when visualized with blue LEDs or Nippon Genetics’ superior Blue/Green LED technology. The second, MIDORI Green Direct, is designed to mix with your sample and load onto a stain-free gel. MIDORI Green Direct has the loading dye included, and detects nucleic acid fragments to a similar, if not slightly better, level as MIDORI Green Advance. The newest member of the family is MIDORI Green Xtra/Easy. Like Advance, Xtra/Easy is added to molten agarose and buffer. It’s chemical structure is optimized for Blue/Green and Blue LED light, leading to unbeatable fluorescence signals of DNA and RNA in agarose gels. MIDORI Green Advance and MIDORI Green Direct dyes are compatible with UV-light, but slightly less efficient when using UV rather than the non-damaging visible excitation light of LEDs. Most importantly for labs, MIDORI Green Xtra/Easy does not stain the agarose gel, leading to an excellent signal-to-noise ratio. Xtra/Easy makes the detection of even the most minute quantities of DNA or RNA possible (and certainly better than EtBr)—it’s not just a replacement for EtBr, it’s an upgrade.

Safer for you and better for your subcloning
By using safe MIDORI Green dyes and safe Blue/Green LED illumination you can improve your subcloning transformation efficiencies by THREE-FOLD. In the example below, a plasmid vector was double digested with suitable restriction enzymes to create two sticky-ended DNA fragments: the lacZ gene (3,536 bp) and the backbone of the vector (4,318 bp). Equal amounts of digested DNA were electrophoresed on 1% agarose gels. The gels were stained with either ethidium bromide or MIDORI Green Direct gel stain according to the corresponding manuals, and then viewed using either a UV transilluminator or the FastGene Blue/Green LED Illuminator, respectively. The two DNA fragments were excised from the gels and purified using a silica membrane based purification kit. The lacZ gene and the vector backbone were re-ligated using T4 DNA ligase transformed into DH5a cells and plated onto selection plates. The total number of blue and white colonies was counted to evaluate cloning efficiency. Each experiment was conducted in triplicate, and the average cloning efficiency was determined. MIDORI Green Direct resulted in a dramatic increase of positive transformants.

Midori Green Can Boost Your Cloning Results!

Ethidium bromide is typically used in conjunction with a strong UV light source to excise DNA bands for purification prior to the ligation reaction. Short-wavelength light is known to cause thymine dimers and damage DNA. The extent of this damage is not always appreciated. High-energy light wreaks havoc on DNA fragments in mere seconds. As can be seen below, cloning efficiency starts to drop after just a 15 second exposure of DNA in a standard agarose gel. After a 30 second exposure, your cloning experiment is all but dead! In contrast, the cloning efficiency of protocols that use blue LEDs or Nippon Genetics’ super-performing Blue/Green LEDs are completely unaffected by this deleterious effect. If your lab can’t to break itself of its ethidium bromide habit, using a Blue/Green LED Illuminator (or imaging system) should still have an immediate positive impact on DNA integrity and cloning efficiency.

UV Transilluminators Kill Cloning Experiments

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