Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

How to avoid gel breakage after electrophoresis


  • Please log in to reply
5 replies to this topic

#1 vivlee

vivlee

    member

  • Active Members
  • Pip
  • 6 posts
0
Neutral

Posted 30 October 2012 - 05:14 PM

Hi all!
I have been running 8% SDS-gel (acrylamide) and the gel often breaks easily when I try to remove it from the glass plate after electrophoresis into a container for staining. I usually keep the gel wet during transferring into the container by letting deionized water running on the plate when I am trying to life the gel, though sometimes succussful, minor breakages still occur at times. Breakages also occurs at time when I need to take the gel out of the container for digital imaging after the staining.
Any SDS-PAGE experts out there that can help me minimize the chance of obtaining a broken gel? Thanks!

#2 John Forsberg

John Forsberg

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 34 posts
13
Good

Posted 30 October 2012 - 07:16 PM

One thing you could try is using a water squirt bottle to soak the gel while it's on one of the plates until an edge lifts up, then trying to use the water stream to kind of wedge the gel into your container for imaging. Or just put the whole plate in with your gel, which might soften up how strongly the gel sticks to your plate, then gently teasing the gel off with a spatula/forceps after incubating/rocking for a bit.

I'm assuming you're using your own gels on glass plates?

#3 vivlee

vivlee

    member

  • Active Members
  • Pip
  • 6 posts
0
Neutral

Posted 30 October 2012 - 07:38 PM

Thanks! I will try loosening the gel edge first with water stream and rock the plate and gel together in water within a container.
And yes, I am using my own gels on glass plates, not commercial gels.

#4 vivlee

vivlee

    member

  • Active Members
  • Pip
  • 6 posts
0
Neutral

Posted 30 October 2012 - 07:53 PM

One thing you could try is using a water squirt bottle to soak the gel while it's on one of the plates until an edge lifts up, then trying to use the water stream to kind of wedge the gel into your container for imaging. Or just put the whole plate in with your gel, which might soften up how strongly the gel sticks to your plate, then gently teasing the gel off with a spatula/forceps after incubating/rocking for a bit.

I'm assuming you're using your own gels on glass plates?


Any tip for removing the gel from water in a container onto the platform in digital imaging instrument without tearing it ?

Edited by vivlee, 30 October 2012 - 07:54 PM.


#5 John Forsberg

John Forsberg

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 34 posts
13
Good

Posted 30 October 2012 - 08:01 PM

Ooh, for imaging, I'll usually have it in pretty deep water/liquid in my tray, then I'll either try to get both (gloved) hands under the edges to lift it out all at once or I'll tease up an edge with some filter forceps, then lift it enough with one hand to get the other hand under it. That way the force of moving it is more distributed and less likely to tear. I wet my gloves before handling gels to help prevent sticking (you should be able to easily slide your finger over the gel without it grabbing).

As for on the imager, I'll usually coat the surface with water first, then set the gel in that one edge first, then easing it down to prevent bubbles.

For getting your gel off the plates, you could also try holding the plate over your container, then teasing a little piece of edge away, then doing the water bottle trick.

#6 vivlee

vivlee

    member

  • Active Members
  • Pip
  • 6 posts
0
Neutral

Posted 30 October 2012 - 09:44 PM

Ooh, for imaging, I'll usually have it in pretty deep water/liquid in my tray, then I'll either try to get both (gloved) hands under the edges to lift it out all at once or I'll tease up an edge with some filter forceps, then lift it enough with one hand to get the other hand under it. That way the force of moving it is more distributed and less likely to tear. I wet my gloves before handling gels to help prevent sticking (you should be able to easily slide your finger over the gel without it grabbing).

As for on the imager, I'll usually coat the surface with water first, then set the gel in that one edge first, then easing it down to prevent bubbles.

For getting your gel off the plates, you could also try holding the plate over your container, then teasing a little piece of edge away, then doing the water bottle trick.


Ok, so i see that the water bottle trick is the key in any transferring. Thanks for the tips! And Happy Halloween! :)




Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.