ligation ratio - (May/23/2007 )
Hi All,
I am trying to clone a 3kb insert into a 4kb vector and not having much luck. Normally for my ligations I use a 5:1 ratio of insert to vector, but am wondering if this applies when the vector and insert are almost the same size! 
Thanks for any help that you guys can offer!
Sarah
-vw3sarah22-
QUOTE (vw3sarah22 @ May 23 2007, 10:15 AM)
Hi All,
I am trying to clone a 3kb insert into a 4kb vector and not having much luck. Normally for my ligations I use a 5:1 ratio of insert to vector, but am wondering if this applies when the vector and insert are almost the same size! Thanks for any help that you guys can offer!
Sarah
I am trying to clone a 3kb insert into a 4kb vector and not having much luck. Normally for my ligations I use a 5:1 ratio of insert to vector, but am wondering if this applies when the vector and insert are almost the same size!
Thanks for any help that you guys can offer!Sarah
A ratio of 1:1 is sufficient for ligation. I routinely do 1:3 or 1:5 and sometimes 1:10 (for difficult cloning).
This applies for all sizes of insert and vector.
-scolix-