Lenti vector ligation - (Sep/16/2011 )
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
-aazmaish-
If you don't get any colonies after ligation, it can be caused by anything from bad ligation to competent cells.
-CloneMan-
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.
-aazmaish-