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
- - - - -

Restriction digest


  • Please log in to reply
1 reply to this topic

#1 dnalab

dnalab

    member

  • Active Members
  • Pip
  • 5 posts
0
Neutral

Posted 03 May 2003 - 09:36 AM

Dear all,
At the moment I have a problem with my restriction digest as the enzyme failed to cleave. I am using PflMI (from NEB) to digest PCR products and used lambda DNA as control. I followed instructions supplied regarding incubation and buffer used. The reaction was supplimented with BSA and left for 18hrs. This enzyme is sensitive to Dam and Dcm methylation. Any suggestions will be appreciated. I was going to decrease the amount of DNA used for digestion, using only 10ul PCR product in a 50ul reaction volume.

Thanks

dnalabmalta@yahoo.co.uk

#2 zhenghj

zhenghj

    member

  • Active Members
  • Pip
  • 8 posts
0
Neutral

Posted 09 May 2003 - 02:09 AM

how about ur control? If u r digesting an enzyme site designed in PCR primer,
it's probably because that ur protection bases outside of the enzyme site is too
short.




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

©1999-2013 Protocol Online, All rights reserved.