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
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2 replies to this topic
#2
CloneMan
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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
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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.
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.
