Ethidium bromide

[Aga]

I found out that EtBr boils at 100 degrees C. This was stated on BioRad product description.
So it seems it may be dangerous when heated and/or added to hot agarose before casting the gel.

[wnym7247]

hi all
this is nice discussion topic.thanks for sharing.
chris

[Warthaug]

I realize I'm late coming to this thread, but I thought I'd share a couple of things we've found in our lab with the syber and etbr dyes.

Firstly, we've found that staining with SYBR seems to interfere with some restriction enzymes and with T4 DNA ligase. As such we've stopped using it for digests/ligations because of the apparent incompatibility. Invitrogen claims SYBR works fine for these applications, but at least in our hands, it seems to interfere.

Secondly, for those worried about disposal, you can (or at least could) get filters that would bind up free EtBr. Simply place over a beaker, pour the solution through, and you get EtBr "free" run-through which can be disposed of the same as any other water-miscible waste. The filter goes to hasmat.

Lastly, we've found that not all equipment setup for EtBr works for SYBR (due to differences in emission wavelength). So while switching may make the lab safer (or at least, gels easier to run), you may need to upgrade some of your equipment.

Bryan

[xsummer]

Originally Posted by Bhappy

Ethidium bromide not a carcinogen in first place....Here i passing you a article which i read long back"

"These in vitro tests, which comprise the entire body of evidence upon which the ethidium bromide hysteria is built, don’t provide any evidence that ethidium bromide can exert a genotoxic effect in anything more complicated than a single cell or an unprotected embryo. In fact there is no direct evidence implicating ethidium bromide as a carcinogen in any animal.

For many years, ethidium bromide has been routinely administered for the treatment of African Sleeping Sickness in cattle. For this purpose, ethidium bromide is administered via subcutaneous or intramuscular injection with no reported increase in incidence of tumor formation or birth defects in the treated cattle. This suggests that ethidium bromide is far less genotoxic to animal systems than is presumed from the in vitro data.

The recommended, apparently non-toxic, dose of ethidium bromide is 1mg/kg of body weight in cattle. In comparison to this, the standard concentration used in molecular biology (around 1 microgram/litre), is low. Rosie Redfield puts it into perspective:

A 50kg researcher would need to drink 50,000 liters of gel-staining solution to get even the non-toxic dose used in cattle.

From this, the risks posed to a scientist handling a very weak solution of ethidium bromide, with a gloved hand (remember the cattle are injected with the stuff) are put into perspective.

A real concern is that the irrational and ill-informed fear of ethidium bromide drives us to solutions that are more dangerous than ethidium bromide itself.

Hello.

that is true but, how about the papers or the evidences of that.

Do you have a link or something like that with evidences?


.Sorry for my english

[grehamjones]

Ethidium bromide is a nucleic acid stain. It fluoresces under ultraviolet light, especially when bound to double-stranded DNA.It is known to be toxic and mutagenic and may be fatal if swallowed, inhaled, or absorbed though the skin.

[balvir_banga]

it is very rightly said that ethidium bromide is toxic when it come in contact with body.But i say any dye which intracts with the DNA is toxic.so they should be handled carefully

[Aga]

Originally Posted by Warthaug

(...) you can (or at least could) get filters that would bind up free EtBr. Simply place over a beaker, pour the solution through, and you get EtBr "free" run-through which can be disposed of the same as any other water-miscible waste. The filter goes to hasmat.

Thanks a lot, maybe you can direct me to the company that produces such filters or a product name? (If you don't wish to give a specific name send it to me via PM, please).
Thanks

[foster033]

Originally Posted by Aga

I found out that EtBr boils at 100 degrees C. This was stated on BioRad product description.
So it seems it may be dangerous when heated and/or added to hot agarose before casting the gel.

I routinely make large quantities of agarose and add EtBr to the melted agarose. Since I make such a large amount I don't use it all at once so I let it solidify in the container I made it in. When I need a gel again I just pop it in the microwave (with/EtBr already dissolved in there) and heat it back up. Never had any issues with this.

Originally Posted by balvir_banga

it is very rightly said that ethidium bromide is toxic when it come in contact with body.But i say any dye which intracts with the DNA is toxic.so they should be handled carefully

EtBr will not absorb directly through your skin. You only need to be worried if you have some sort of cut or abrasion in which it could enter through. Also, EtBr can seep through latex gloves, so you should always wear nitrile gloves when handling EtBr if you want your skin to be protected.

[dlovan]

for gel electrophoresis(1gm agarose+100 ml TBE buffer) after boiling ? the exact concentration of ethidium bromide will be?
note if we have stock soln. of etbr(500microgram/ml).

[Sharda Bhagchandani]

2microgram in 100ml is sufficient enough for detecting bands. However, for safer side use 4 microgram instead of two.
So from your stock solution, you can use 8 microliter