Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > General Forum > Article Discussion
Register Search Today's Posts Mark Forums Read

Article Discussion Discuss Molecular Biology Topics from the Molecular Biology Wiki. Article Discussion Forum.


Ethidium bromide

Ethidium bromide - Article Discussion

Ethidium bromide - Discuss Molecular Biology Topics from the Molecular Biology Wiki. Article Discussion Forum.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 06-10-2007, 10:32 AM
admin's Avatar
Administrator
 
Join Date: Nov 2005
Posts: 1,418
Thanks: 883
Thanked 68 Times in 58 Posts
Default Ethidium bromide



Discussion thread for Ethidium bromide. If you would like to add a comment, click the Post Reply button.
Reply With Quote
The Following User Says Thank You to admin For This Useful Post:
wnym7247 (09-27-2009)
  #2  
Old 12-29-2008, 08:53 AM
Pipette Filler
Points: 348, Level: 6 Points: 348, Level: 6 Points: 348, Level: 6
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2008
Location: Sydney
Posts: 8
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Ethidium bromide

Its a very powerful mutagen so MSDS should be read before handling it.
Reply With Quote
  #3  
Old 12-30-2008, 10:25 AM
Aga's Avatar
Aga Aga is offline
Post-Doc
Points: 2,239, Level: 30 Points: 2,239, Level: 30 Points: 2,239, Level: 30
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Nov 2008
Posts: 270
Thanks: 25
Thanked 78 Times in 64 Posts
Default Re: Ethidium bromide

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?
Reply With Quote
  #4  
Old 12-31-2008, 08:53 AM
admin's Avatar
Administrator
 
Join Date: Nov 2005
Posts: 1,418
Thanks: 883
Thanked 68 Times in 58 Posts
Default Re: Ethidium bromide

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
Reply With Quote
  #5  
Old 01-02-2009, 05:56 PM
Aga's Avatar
Aga Aga is offline
Post-Doc
Points: 2,239, Level: 30 Points: 2,239, Level: 30 Points: 2,239, Level: 30
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Nov 2008
Posts: 270
Thanks: 25
Thanked 78 Times in 64 Posts
Default Re: Ethidium bromide

Quote:
Originally Posted by admin View Post
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?
Reply With Quote
  #6  
Old 01-04-2009, 12:17 PM
Aga's Avatar
Aga Aga is offline
Post-Doc
Points: 2,239, Level: 30 Points: 2,239, Level: 30 Points: 2,239, Level: 30
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Nov 2008
Posts: 270
Thanks: 25
Thanked 78 Times in 64 Posts
Default Re: Ethidium bromide

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?
Reply With Quote
The Following User Says Thank You to Aga For This Useful Post:
admin (01-17-2009)
  #7  
Old 01-17-2009, 10:58 AM
admin's Avatar
Administrator
 
Join Date: Nov 2005
Posts: 1,418
Thanks: 883
Thanked 68 Times in 58 Posts
Default Re: Ethidium bromide

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
Reply With Quote
  #8  
Old 01-20-2009, 05:12 PM
Pipette Filler
Points: 1,764, Level: 25 Points: 1,764, Level: 25 Points: 1,764, Level: 25
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Dec 2008
Location: India
Posts: 4
Thanks: 0
Thanked 1 Time in 1 Post
Default Re: Ethidium bromide

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.
Reply With Quote
  #9  
Old 01-20-2009, 05:54 PM
butters's Avatar
M.D/Ph.D
Points: 4,938, Level: 47 Points: 4,938, Level: 47 Points: 4,938, Level: 47
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Dec 2008
Location: Malaysia
Posts: 315
Thanks: 25
Thanked 68 Times in 63 Posts
Default Re: Ethidium bromide

Quote:
Originally Posted by Aga View Post
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

Quote:
Originally Posted by Aga View Post
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.

Last edited by butters; 01-20-2009 at 05:56 PM.
Reply With Quote
  #10  
Old 01-20-2009, 06:31 PM
Aga's Avatar
Aga Aga is offline
Post-Doc
Points: 2,239, Level: 30 Points: 2,239, Level: 30 Points: 2,239, Level: 30
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Nov 2008
Posts: 270
Thanks: 25
Thanked 78 Times in 64 Posts
Default Re: Ethidium bromide

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?
Reply With Quote
Reply

Tags
bromide , ethidium


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
ethidium bromide contamination on clothes Jayakumar, R Protocols and Methods Forum 12 07-14-2010 05:01 PM
Removal of Ethidium Bromide - Disposal of Ethidium Bromide kiki06 Basic Lab Protocols and Techniques 10 12-29-2009 01:17 PM
Strong contamination with ethidium bromide Undergrad Protocols and Methods Forum 6 06-14-2007 03:12 PM
ethidium bromide vapour? jg374@cam.ac.uk Protocols and Methods Forum 4 09-27-2005 04:00 PM
ethidium bromide vapour? Jayakumar, R Protocols and Methods Forum 1 09-14-2005 02:42 PM


All times are GMT. The time now is 09:12 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.17660 seconds with 16 queries