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
- - - - -

Lenti vector ligation


  • Please log in to reply
2 replies to this topic

#1 aazmaish

aazmaish

    member

  • Active Members
  • Pip
  • 8 posts
0
Neutral

Posted 16 September 2011 - 07:31 AM

Hi,
I am new to this.I need help.
My lenti vector is 7.5kb and insert is 4.5kb. I did ligations but i could not get colonies after transformation. Kindly someone help me in this regard? Please Please.

Regards

#2 CloneMan

CloneMan

    member

  • Active Members
  • Pip
  • 8 posts
0
Neutral

Posted 17 September 2011 - 07:23 AM

If you don't get any colonies after ligation, it can be caused by anything from bad ligation to competent cells.

#3 aazmaish

aazmaish

    member

  • Active Members
  • Pip
  • 8 posts
0
Neutral

Posted 17 September 2011 - 08:30 AM

Hi CloneMan,
Thanks a lot.
I used the same ligation protocol which i always used for other ligations, (that is T4 ligase), and always i got very good results. The only difference at this time is the insert and vector size. I never worked with 4.5 kb insert and 7.5 kb lenti vector. Would you suggest me some protocol to solve this problem.
Thanks in advance.




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

©1999-2013 Protocol Online, All rights reserved.