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
- - - - -

Using SDS for DNA electrophoresis


  • Please log in to reply
3 replies to this topic

#1 Raygoza

Raygoza

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 50 posts
0
Neutral

Posted 07 September 2015 - 04:48 AM

Now, the question title may sound weird right off the bat, but my question is as follows:

 

Some restriction enzymes warn about the DNA mobility changing properties of some enzymes because the enzyme may remain attached to the DNA, so my question is can i add SDS to the loading dye and perhaps resolve this problem to detach proteins off the DNA ?  i wanted to make sure that SDS doesn't have DNA denaturing properties as well. or if the idea sounds reasonable.

 

I know that you may inactivate some of the enzymes before electrophoresis but please focus on my question as it is.

 

Thanks


Edited by Raygoza, 07 September 2015 - 04:49 AM.


#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 6,722 posts
567
Excellent

Posted 07 September 2015 - 12:12 PM

I think for SDS to work you would need to heat the sample to about 70 C, which would probably denature (some) DNA and might result in further migration problems.



#3 hobglobin

hobglobin

    Growing old is mandatory, growing up is optional...

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,615 posts
127
Excellent

Posted 07 September 2015 - 12:57 PM

NEB suggests to add 0.5.-1 % SDS to the loading buffer to dissociate the enzyme from the DNA when you have such problems, therefore it should work.
DNA will not be denatured except you will heat it or add chemicals such as formamide or dimethyl sulfoxide. The mechanism and result of denaturation in DNA is quite different compared to proteins....and in DNA usually reversible.



A single lie is reproachable; a million lies is a statistic.
D. J. T.


#4 Andrea Fortina

Andrea Fortina

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 55 posts
1
Neutral

Posted 21 June 2016 - 06:17 PM

I saw DNA loading buffer from Thermo contains 1% SDS, which could eliminate DNA-protein interactions, prevent appearance of additional bands due to annealing of DNA molecules with cohesive ends.


Andrea Fortina

Genius only means hard-working for all one's life.

CreativeBiomart, Recombinant protein expert

 

 





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

©1999-2013 Protocol Online, All rights reserved.