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
- - - - -

homolog detection using Southern


  • Please log in to reply
1 reply to this topic

#1 biobutterfly

biobutterfly

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 01 March 2010 - 06:44 PM

I am trying to use a Southern to detect the number of homologs. I suspect there are three but need to test. If I use a dot blot to determine the concentration of gDNA to run out on the gel, will that number actually be correct? If, for example, my probe shows a nice signal with 10ug dotted, won't the signal be much weaker when the DNA is all separated out on the gel? Is that linear? Can I just scale up?

Thanks for the help.

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,618 posts
387
Excellent

Posted 03 March 2010 - 03:43 PM

If you are using all three probes in the same hybridisation, the the signal for each should be approximately a third of what you see on the dot blot, assuming equal binding and labeling of the probes.

I would try to dot blot with each of the probes separately to see the signal strength of each, and then use the weakest as the basis for loading.




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

©1999-2013 Protocol Online, All rights reserved.