MIDORIGreen Advance is a safe alternative to the traditional nucleic acid stain ethidium bromide. It is a non-carcinogenic and less mutagenic dye for detecting dsDNA, ssDNA and RNA in agarose gels with a very high sensitivity. MIDORIGreen Advance can utilize with UV light or with our innovative Blue/Green LED technology.
The next generation of in-gel staining
MIDORIGreen Advance is a second-generation non-carcinogenic dye. Optimised for a brighter signal when excited by UV-light or blue/green light. It maintained the advantages, such as being non-carcinogenic and having an excellent noise-to-signal ratio. As seen in Fig. 1 MIDORIGreen Advance is even better than ethidium bromide when using the Blue/Green LED technology.
Fig. 1: Comparison of sensitivity between MIDORIGreen Advance (left green) and ethidium Bromide (right red) using a Blue/Green LED Transilluminator.
MIDORIGreen Advance shows a very high sensitivity even for small DNA fragments. The dilution factor of MIDORIGreen Advance can be as high as 1:25000. Hence, 4-6 μl are enough for the staining of 100 ml agarose gel, resulting in ~17 to 25 liters of stained agarose gels.
It is essential for a good replacement of the mutagenic DNA stain ethidium Bromide to deliver strong signals. MIDORIGreen Advance delivers signals with a comparable intensity. Nonetheless, the safety of the user must not be compromised. Hence, several tests were performed with MIDORIGreen Advance and according to those tests, MIDORIGreen Advance is safe.
Download here the MIDORIGreen Advance safety report.
More than 500 citations in journals!
MIDORIGreen Advance has been used very successfully in many laboratories. The feedback from the scientific society was very positiv. Enclosed a few results from independent scientists:
gDNA was extracted from rapeseed and sunflower seed using the Clean Plant PK Kit
“…we have tested Midori Green Advance and we are very excited about it. Please find attached to this email a typical picture….”
Dr. Olaf Voolstra,
Biosensor Research Group, Hohenheim University Stuttgart, Germany
Ethidium bromide (left) in comparison with MIDORIGreen Advance (right) on UV table
Data kindly provided by
Friedrich-Alexander-University Erlangen-Nürnberg, Germany
Comparison of ethidium bromide and MIDORIGreen Advance on a UV table
Data kindly provided by
Heinrich-Heine-University Düsseldorf, Germany
- Prepare 100 ml of agarose gel solution (concentration from 0.8-3.0%) and heat until the solution is completely clear and no small floating particles are visible.
- Add 4-6 µl of MIDORIGreen Advance DNA Stain to the gel solution and mix it gently. Cool the gel to 50- 60ºC and cast the gel, into the gel tray. When the gel is solid, load the samples and perform electrophoresis.
- Detect the bands using a UV or LED illuminator. Yellow or green gelatin- or cellophane filters should be used for photography.
- MIDORIGreen Advance DNA Stain poststaining solution may be used 2-3 times.
- Staining solution that is going to be reused should be preferably stored at room temperature in the dark.
- For <0.5 cm thick agarose gel, 10-25 µl of the stain should be used per 100 ml of buffer.
- Optimal staining time (5 – 60 minutes) and the amount of the stain may depend on the thickness of the gel.
NOTICE: Usage is not recommended with SDS containing loading buffers because of band appearance caused by stain and SDS interaction!
Can I use Midori Green Advance with Pulse Field Gel Electrophoresis (PFGE)? Normaly I used a poststaining with EtBr.
A poststaining is done with MGA by protocol which you can find in the protocol section.
Do you have any details about the disposal of Midori Green Advance please? I understand that the molecule is light-sensitive and I wondered if the MGA-gel could be exposed to light after use before disposal?
MGA is not toxic and it is sufficient to dispose it as chemical lab waste. The emissivity of MGA will be destroyed by light, but that does not mean that the chemical structure will be changed. But it is not necessary to destroy the molecule because it is anyway not harmful. That is the big advantage in comparison to ethidium bromide – that there is no special disposal necessary.
Why can’t MGA and MGD be used the same way?
The chemical structures are completely different. we tested to use MGD as MGA and vice versa. We can’t recommend both applications. As you try to use MGD as MGA, you need a lot of MGD and ladder in order to get observable bands. Samples with a low concentration could be stay undetected by this way. MGD was designed for addition to sample and is much less concentrated than MGA. It is very uneconomic to use MGD in the way like MGA – you need more than the 10 fold amount in the gel in order to observe bands. As you try to use MGA as MGD it is also problematic, because MGA is too high concentrated and is running in the opposite direction. I tried to reduce the MGA concentration, in this case you see some bands, but they are very weak independent which dilution is used. Summarizing you can say both stains are designed (with different structural formula) and optimized for different applications (in gel and addition to a sample) and it is not recommended to try to use them in another way.
What is the difference between MGA and MGD and how stable are these stains?
The main difference is that Midori Green Advance (MGA) is for in gel and post staining (means it is used like ethidium bromide) and Midori Green Direct (MGD) is added directly to the sample. Depending on the gel documentation system we recommend to use one or the other but you can use both for every illuminator (UV 302, 365 nm, Blue illuminator and Blue/Green transilluminator) – only the performance can be increased to an optimal level. For example we recommend to use MGA if the user has a UV transilluminator, because the results are fantastic with that stain. A really great performance you get with our Blue/Green LED instruments and MGD – no background at all and an intensive signal. Both stains are stable on room temperature, so shipping at RT is no problem, but we recommend to store the stains at 4°C.
Are MGA and MGD working with Agarose and acrylamide gels?
Our customers gave us the feedback that MDG and MGA (post staining) are working for acrylamide gels but primarily they are designed for agarose gels.
What happens if I stored MGA at -20°C for 6 days? Will it be OK for use?
The freezing destroys the functionality of MGA. You can still try but our experience it forfeits the illumination after freezing.
I use Midori Green Advance in Agarose Gels, why are the small bands invisible when I run a gel for 35 minutes and what do you recommend?
Midori Green Advance is possitively charged and runs in the different direction than the DNA, therfore the gel gets destained from the bottom. To see also small bands you can reduce the running time to 25 minutes. If you want to run your gel for a longer time, than you can post stain. You can use the in gel staining and then have a shorter time for post staining or you only post stain your gel without a previous in gel staining. In both cases the whole gel will be stained and also very low molecular weight bands are observable. You can also use Midori Green Direct instead of Midori Green Advance. This stain is added to the DNA so every band is stained no matter how long you run the gel. Another advantage is the nonexistent background. In special if you use blue or Blue/Green LED light instead of UV, you will get amazing signals.
On the Midori Green Advance Datasheet it is described that postaining solution may be used up to 2-3 times. For how long could you use the postaining solution without losing too much sensibility?
Actually, we have customers who use it more often than 2-3 times. If the staining bath is darkened the whole time, you can use it in any case 2-3 times without any reduction of signal intensity. But it is also possible to use it 5 – 8 times without a signal decrease depending on the size of the gel and the amount of samples. If you detect a decrease of intensity, it is possible to add a little bit MGA and thus get same signal intensity as before.
Can I use MGA after expiry date?
Yes, you can use it. The expiry date is just the minimal amount of time we guarantee but normally it last much longer.
I get some additional bands after using MGA?
This is possible, if you use SDS in the loading buffer.
Can you use Midori Green in a Polyacrylamid/Urea Gel?
This works really nice.
Is Midori working in Urea/Formamide gels?
Unfortunately, it is not working. That’s because of the Formamide. Urea gels are working.
Does Midori Green contain DMSO?
Our Midoris are DMSO free.
Can I use marker containing Orange G, Bromphenol blue or Xylen cyanol FF with Midori Green Advance?
Yes, you can use them. They do not cause probles, only Bromphenol blue can reduce the signal intensity or can lead to formation of clouds in the gel.
Can I add Midori Green Advance to the running buffer to reduce the destaining of the gel? If yes in which amount should I use it?
You can use Midori Green Advance in the running buffer. Here you should use arond 8 µL per 100 ml of buffer.
Can I use Midori Green Advance with further downstream applications like southern blots?
Customers tell us that there the dye does not interfere with standard Southern-protocols. We have no testing at our side, since there is no indication that the dye intherferes with procedures downstream of agarose gel-runs and – documentation.The only (indirectly) related issue is certainly, that no direct comparison can be drawn between Southern-blots that have been stained in-gel (during the electrophoresis) with EtBr in first experiments and later experiments that employed MG advance, since gel-run patterns are not directly comparable between these two dyes (if used in-gel). For any post-run staining and down-stream application there are no known issues.
Can Midori Green Advance be used in denaturating Formaldehyde Gels for separating RNA?
Yes, it can be used:
We have separated total RNA from 1% Formaldehyde gel with TAE buffer using both ways Post staining and in Gel Staining. In each case we used 5 microliters per 100 ml buffer. Post staining was carried out at room temperature for 1 hour.
The minimum RNA concentration should be at least 250 ng per line. The good working RNA concentration in 500 ng.