MIDORI Green Frequently Asked Questions Can I use Midori Green Advance with pulse field gel electrophoresis (PFGE)? Normally I used a poststaining with EtBr.
A post-consultation is done with MGA per protocol, which can be found in the protocols section.
Do you have any details about Midori Green Advance removal please?
I understand that the molecule is sensitive to light and I wondered if the MGA gel could be exposed to light after use before disposal
MGA is not toxic and is sufficient to dispose of it as chemical laboratory 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 not harmful anyway. That is the great advantage compared to ethidium bromide: that no special elimination is necessary.
Why can’t MGA and MGD be used in the same way?
The chemical structures are completely different.
We tested the use of MGD as MGA and vice versa. We cannot recommend both applications. As you try to use MGD as an MGA, you need a lot of MGD and ladder to get observable bands. Samples with a low concentration may not be detected in this way. MGD was designed to be added to the sample and is much less concentrated than MGA.
It is very unprofitable to use MGD in the same way as MGA: it takes more than 10 times the amount in the gel to observe the bands. When you try to use MGA as MGD, it is also problematic, because MGA is too concentrated and is running in the opposite direction. I tried to reduce the concentration of MGA, in this case you see some bands, but they are very weak, regardless of the dilution used. In summary, you can say that both spots are designed (with a different structural formula) and optimized for different applications (in gel and in 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 spots?
The main difference is that Midori Green Advance (MGA) is for gel and post staining (means that it is used as ethidium bromide) and Midori Green
Direct (MGD) is added directly to the sample. Depending on the gel documentation system, we recommend using one or the other, but you can use both for each illuminator (UV 302, 365 nm, blue illuminator and blue / green transilluminator):
you can only increase the performance to an optimal level. For example, we recommend using MGA if the user has a UV transilluminator, because the results are fantastic with that staining. Really excellent performance you get with our blue / green and MGD LED instruments, with no background at all and an intense signal. Both stains are stable at room temperature, so shipping at room temperature is not a problem, but we recommend staining them at 4 ° C.
Do MGA and MGD work with agarose and acrylamide gels?
Our clients informed us that MDG and MGA (subsequent staining) are working for acrylamide gels, but they are primarily designed for agarose gels.
What happens if I stored MGA at -20 ° C for 6 days? Will it be fine for use?
Freezing destroys the functionality of MGA. You can still try, but our experience loses the lighting after freezing.
I use Midori Green Advance in Agarose Gels, why are small bands invisible when I run a gel for 35 minutes and what do you recommend? Midori Green Advance is positively charged and runs in a different direction than the DNA, so the gel is destroyed at the bottom.
To see also small bands you can reduce the operating time to 25 minutes. If you want to run your gel for longer, you can publish the stain. You can use gel staining and then have a shorter time for subsequent staining or just stain your gel without prior gel staining. In both cases, the entire gel will be stained and bands of very low molecular weight will also be observed. You can also use Midori
Green Direct instead of Midori Green Advance. This staining is added to the DNA so that all bands are stained regardless of how long the gel runs. Another advantage is the nonexistent fund. Especially, if you use blue or blue / green LED light instead of UV, you will get incredible signals.
In the Midori Green Advance data sheet, it is described that the postain solution can be used up to 2-3 times. How long could I use the postain solution without losing too much sensitivity?
Actually, we have customers who use it more than 2 to 3 times. If the staining bath darkens all the time, you can use it in any case 2-3 times without any reduction in signal strength. But it is also possible to use it 5 to 8 times without a decrease in the signal, depending on the size of the gel and the number of samples. If it detects a decrease in intensity, it is possible to add some MGA and get the same signal strength as before.
Can I use MGA after the expiration date? Yes, you can use it. The expiration date is only the minimum amount of time we guarantee, but it usually lasts much longer.
Do I get some additional bands after using MGA?
This is possible, if you use SDS in the load buffer.
Can Midori Green be used on a polyacrylamide / urea gel?
This works very well. Is Midori working on urea / formamide gels? Unfortunately, it is not working. That is due to formamide. Urea gels are working.
Does Midori Green contain DMSO?
Our Midoris are free of DMSO. Can I use ladders that contain Orange G, Bromphenol blue or Xylen cyanol FF with
Midori Green Advance?
Yes, you can use them. They do not cause problems, only bromphenol blue can reduce the intensity of the signal or can lead to cloud formation in the gel.
Can I add Midori Green Advance to the execution buffer to reduce gel deterioration? If yes, how much should I use it?
You can use Midori Green Advance in the running buffer. Here you should use about 8 µL per 100 ml of buffer.
Can I use Midori Green Advance with other later applications such as southern transfers?
Clients tell us that there the dye does not interfere with the standard protocols of the South. We have no evidence at our side, as there is no evidence that the dye interferes with the downstream procedures of the agarose gel and the documentation.
The only related problem (indirectly) is certainly that no direct comparison can be made between Southern blots that have been stained in gel (during electrophoresis) with EtBr in the first experiments and in subsequent experiments that employed the advance of MG, since the gel performance patterns are not directly comparable between these two dyes (if used in gel). For any post staining and downflow application there are no known problems.