Dear Fatima,

EtBr is usually added to agarose solution, once it is cool. You dont need to add it during boiling or in very hot solution. boil agarose in TAE buffer and then cool it. you can do cooling under tap water for 2-3 min and when it is warm just add EtBr mix it swirling to avoid air bubbles and pour it immediately. EtBr molecular weight is 394 so quite large molecule and melting temp is >250. so the chances for evaporation are very less. Very few molecules can degrade during boiling that may come into vapours.

wear couple of gloves in each hand ..after you are done, wipe the surface wit 70% ethanol. Best way is you have a specified place in lab to do this. but if you dont have then just clean the area after you are done. and dispose off properly.

you dont need safety cabinet for EtBr. In most of the laboratories, agarose gel system in usually separated from other benches to prevent spill over contamination and avoid contact. you should be care full about the gloves you use for handling the agarose gels, buffers. some of the gloves are not suffieciently resistant for this purpose, so read the instructions, or ask from the manufacturers. As Ranjan suggested you should be carefull is disposing off EtBr containing liquids and gels.

thank u for ur response.if u dont mind pls clarify my these doubts also.while adding EtBr into hot agarose in conical flask definitely vapoures will come out.the inhalation of that vapours containing EtBr also dangerous.so to prevent that is it necessary to use safety cabinet/laminar flow?

Add EtBr in running buffer rather than in hot agarose. There will be no vapours then and you will be on safer side.

Dear Fatima,

EtBr is usually added to agarose solution, once it is cool. You dont need to add it during boiling or in very hot solution. boil agarose in TAE buffer and then cool it. you can do cooling under tap water for 2-3 min and when it is warm just add EtBr mix it swirling to avoid air bubbles and pour it immediately. EtBr molecular weight is 394 so quite large molecule and melting temp is >250. so the chances for evaporation are very less. Very few molecules can degrade during boiling that may come into vapours.

protect ur hands well

Thank u to all responded to me.

hi Birendra Singh,

thanks a lot for ur detailed reply.

You don't have to use a safety cabinet, plus please make sure that you wear proper gloves when handling EtBr and try to work on a separate bench, away from others.

if you expose ethbro to light/air a bit longer, it will be degarded, keep the solution in dark and direct skin contact could be cancerous.

is EtBr inhalation is dangerous?

of course it is.

never put EtBr. in hot agarose. Let it cool a little and then add Et.Br. Keep thesolution in a dark bottle with almunium foil wraped arround the bottle. Avoid direct exposure to light and heat.

Avoid to use EtBr in the solid / pouder form. If possible always buy it as a solution (ready to use) so there is no need to weigh it in a safety cabinet. Inhaling EtBr as a pouder is the worst you can do to your body. When handlich solutions containing EtBr allways wear gloves.

Add EtBr to the agarose solution when the heat comes down a little before slab preparation,load the sample,run and view.this is easier and safer than going for staining after running the gel.

is it necessary to use atleast a fume hood while working with EtBr?

You can work with EtBr on the bench. You have to make sure that you set up a separate area for yourself and that you don't contaminate any other instruments/tools with EtBr. Also make sure that nobody touches the solution and the tools that have been contaminated with it. Use proper gloves for protection, do not allow contact with the skin.

Dear Fathima, first of all, wish you good luck for your research work. I understand the concern you have over using EtBr and appreciate the way people have extended their help. In not-so-rich labs like the one you might have at your University (I have some idea about Indian labs since I come from India), it is difficult to afford separate room/fume hood/special infrastructure. So, we need to manage with existing facilities. You already got some valuable advice from other members. Please also remember to use proper eye protection (protective goggles) beside using gel protection cover (specially if your work involves excising bands from the gels). Preferably try to use EtBr in a staining solution (you can use it from a stock solution too...to avoid preparing fresh solution every-time you run a gel) after removing the gel from your running buffer, rather than adding EtBr to your gel (as suggested by Ms. Shukla). Ten minutes of staining time is enough. You won't have to worry about vapor, etc. in this case.

Fathima, when I was working with EtBr in Brazil, we used to place the gel, after electroforesis, on a EtBr solution, for few minutes. That will prevent your exposure to it and will dye your gel as well. And also, we considered all the utensils used to do it, contaminated. The less you get in touch with it, the better, and remember to discard it appropriately.

thank u very much Mr.Jayashanker

You are most welcome....All the very best for your future....Cheers!