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!
- - - - -

Low blotting efficiency (southern blotting)

  • Please log in to reply
No replies to this topic

#1 TCT



  • Members
  • Pip
  • 2 posts

Posted 31 March 2005 - 04:26 AM


I have a problem with getting all of my DNA (genomic and plasmid mix) transfered from my 0.7% agarose gel (SeaKem LE) to my membrane (Nytran N membrane, Schleicher & Schuell...)

The total DNA is digested with Hind III before electroforesis, and the gel is run at 80 V for 3 hours. The gel has been depurinated with 0,25M HCl in about 10 min, before denaturation and neutralization. The blotting procedure are performed with a turbo blotting system and with 20x SSC as transfer buffer. Blotting time that has been tried out is 12 hours, 46 hours.
12 hours gives a poor transfer, while 46h gives a bit better transfer. How can I increase the transfer?

I have tried a alkaline transfer, but that doesn`t give any better result.

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

©1999-2013 Protocol Online, All rights reserved.