Dismiss Notice
Join Physics Forums Today!
The friendliest, high quality science and math community on the planet! Everyone who loves science is here!

Help with gel electrophoresis

  1. May 30, 2013 #1
    I am a biophysics grad student at Michigan State, and the lab I am working in has just started making 1% agarose gels for DNA purification. The problem is when we try and remove the comb from the gel and the gel from the walls of the form, it sticks to the sides, ripping the gel and rendering it unusable. The protocol that we are following as worked for my PI in other labs, but never for myself (or my PI) in this lab. All our equipment, chemicals, and glassware are new because this is a new lab.

    So, have any of you had a similar problem? What do you think could be causing this?

    I have made sure that it is not the agarose or TBE buffer that is contaminated.
    Here is the protocol:
    Add 0.6g of low melting point LMP Agarose (UltraPure brand) to 60mL of 0.5x TBE buffer (Fisher brand). Microwave until just boiling and mix until all the agarose is dissolved. Allow to cool, add 1μL of EtBr and pour into mold. Wait ~1 hour and remove gel from mold.

    Like I said, my PI used this protocol all the time at his past lab, and he never had this problem.
    Thanks so much for your help.
     
  2. jcsd
  3. May 31, 2013 #2
    Have you tried letting it sit for a little longer? The only other thing I can think of is putting it in the fridge for the last 15 min. or so.
     
  4. May 31, 2013 #3

    Monique

    User Avatar
    Staff Emeritus
    Science Advisor
    Gold Member

    I made hundreds of 1% gels, also on new equipment, and have never had that happen. Be gentle when removing the comb, it helps to start the lifting from one side so that air can come into it. Otherwise contact the company for advice.
     
Know someone interested in this topic? Share this thread via Reddit, Google+, Twitter, or Facebook




Similar Discussions: Help with gel electrophoresis
  1. Gel Electrophoresis (Replies: 5)

  2. CHEF gel trouble (Replies: 3)

Loading...