Ethidium bromide

[admin]

Discussion thread for Ethidium bromide. If you would like to add a comment, click the Post Reply button.

[Nasiara]

Its a very powerful mutagen so MSDS should be read before handling it.

[Aga]

Has anyone tried to get rid of ethidium bromide in his lab? SYBR dyes are the alternative.

We use ethidium bromide from time to time. I hate it having in mind it's carcinogenic, so I turned to real-time PCR as quickly as the opportunity came. Hovewer I will have to use it for other purposes than electrophoresis, sadly...

BTW how do you use ethidium bromide in your electrophoresis?

- Do you incorporate the dye in a gel before casting and perform the electrophoresis in a gel with a dye in it

- or do you use it's solution after the electrophoresis to soak the gel to get the results visualized?

[admin]

Hello Aga,
those are all good points.

The best way to visualize and keep things safe (and easy disposal) is to incorporate the ethidium bromide inside the gel (after the microwaved gel cools a bit).

The problem is, many people that use/buy the imaging equipment dont want ethidium bromide gels on their surfaces (or keyboards).

So the way to go is SYBR dyes which are a bit more expensive or to buy a dedicated ethidium bromide camera system.

We would clean up spills or dirty contaminated areas with activated charcoal and then into ethidium bromide specific disposal containers never into the sinks or regular garbage.

Happy New Year!
Cheers

[Aga]

Originally Posted by admin

The best way to visualize and keep things safe (and easy disposal) is to incorporate the ethidium bromide inside the gel (after the microwaved gel cools a bit).

That's the way we do it and because we always have done it this way I have never seen the difference in visualization of electrophoretic results.
Once I read that ethidium bromide incorporated into the gel prior to electrophoresis gives less sharp/less visible bands ,or something like that, comparing to staining the gel after electrophoresis in ethidium bromide solution.

So, apart the differences in handling, safety and disposal, what's the difference between the two staining approaches in results obtained?
I've watched many vids where the gel is stained with ethidium bromide after elecrophoresis - I consider it less safe. If it is so, why is the method used? What are its advantages?

[Aga]

I don't think any of the posts of hari001 has made this discussion more interesting or informative so far. Quite the opposite.

Is this usually tolerated?

[admin]

nope he was a spammer trying to get more than 10 posts so he can post links. Funny actually, as even his link/post was moderated (in queue) after all that work....thanks to your help I deleted his posts and banned him.

Sorry we can't be on here 24-7, thats why we have some automated protections (many actually email banning, bot banning, spam keywords, img/link/url protection) but we are always looking for more moderators if you are interested

[Sharda Bhagchandani]

EtBr, a potent mutagen, has great significance in DNA electrophoresis as it omits out use of any staining. However its high conc can affect DNA quality and its visuality. I think, a good discussion on it is possible.

[butters]

Originally Posted by Aga

Has anyone tried to get rid of ethidium bromide in his lab?

BTW how do you use ethidium bromide in your electrophoresis?

- Do you incorporate the dye in a gel before casting and perform the electrophoresis in a gel with a dye in it

- or do you use it's solution after the electrophoresis to soak the gel to get the results visualized?

I love ethidium bromide!!! It always remind me to handle all chemical in lab with upmost precaution & respect. my etbr is usually in pre stain gel

Originally Posted by Aga

That's the way we do it and because we always have done it this way I have never seen the difference in visualization of electrophoretic results.
Once I read that ethidium bromide incorporated into the gel prior to electrophoresis gives less sharp/less visible bands ,or something like that, comparing to staining the gel after electrophoresis in ethidium bromide solution.

So, apart the differences in handling, safety and disposal, what's the difference between the two staining approaches in results obtained?
I've watched many vids where the gel is stained with ethidium bromide after elecrophoresis - I consider it less safe. If it is so, why is the method used? What are its advantages?

pre-stain gel(mix with etbr before solidified)
advantages
fast, no need destain, generally save time and easy to prepare
con
not good for DNA quantification, especially the lower base pair DNA

** reason
DNA more from (-) to (+) [correct me as i sometime mix up] however etbr move the opposite direction. (+) to (-). thus when time goes by there will be less etbr in the lower end / low base pair region. this result in lower intensity and not accurately reflective of the concentration.

[Aga]

Yeah, we pre-stain gels for similar reasons.

Disadvantages - good remark Butters. That's fortunate we don't need to use this method for DNA quantification.

My question - is ethidium bromide volatile? I mean it depends on the temperature, but what's the boiling point? Is it very volatile at 50-60 degrees C?