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
- - - - -

gene design


  • Please log in to reply
2 replies to this topic

#1 roshanbernard

roshanbernard

    member

  • Active Members
  • Pip
  • 26 posts
0
Neutral

Posted 10 April 2009 - 12:20 AM

hi,
i am designing a synthetic gene and i need to have Sal1and EcoRV restriction site at the 5' and 3' terminals.can i just insert the restriciton site of the enzyme on both ends and then design the whole gene..or do i need any additional base pairs in addition to these restriction sites on 5' and 3' ends....

#2 phage434

phage434

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 2,504 posts
252
Excellent

Posted 10 April 2009 - 03:57 AM

You need additional bases. I would add at least 6, preferably 8 bases 5' of your cut site. I would suggest switching to a different enzyme than SalI, which has a very poor reputation for cutting PCR fragments.

#3 perneseblue

perneseblue

    Unlimited ligation works!

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 578 posts
17
Good

Posted 10 April 2009 - 10:05 PM

I would suggest switching to a different enzyme than SalI, which has a very poor reputation for cutting PCR fragments.


XhoI and SalI form compatible overhangs
May your PCR products be long, your protocols short and your boss on holiday




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

©1999-2013 Protocol Online, All rights reserved.