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
- - - - -

gDNA restriction digest


  • Please log in to reply
1 reply to this topic

#1 proteinz

proteinz

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 36 posts
0
Neutral

Posted 08 October 2010 - 07:44 AM

Hi All,
I'm a bit confused. I'm attempting to perform a partial restriction digest of gDNA. but What should this look like on a gel? i want bands between 1kb-12kb. Should it just be a a smear in my size range? And Should there be undigested gDNA visible? Also what would a complete digestion of the gDNA look like?I am restricting with EcoRI
Many thanks for your time!

#2 hobglobin

hobglobin

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

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,465 posts
90
Excellent

Posted 08 October 2010 - 09:02 AM

It depends on DNA (i.e. species) and the enzymes you use. EcoRI is normally a frequent cutter (i.e., the recognition site has six base pairs frequently occurring in the genome) and will produce a lot of smaller fragments, resulting in a smear on the gel. If you use enzymes whose recognition sequences are less frequent in your DNA, then you'll get larger fragments, and also less (some enzymes might even not cut at all).
Normally you have to try out which enzyme(s) produce the range of fragment sizes you need. Or you find a publication with some information about this.

Edited by hobglobin, 08 October 2010 - 09:18 AM.

One must presume that long and short arguments contribute to the same end. - Epicurus
...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.




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

©1999-2013 Protocol Online, All rights reserved.