TA Clonning

[Scott]

> I tried to clone a PCR product. However, after transform the Top10
> compentence cells, I did not get clones. The PCR is good. I phosphoralted
> the 5' end of the primer. I used Invitragen Company's kits. Ligation
> condition is at 16 degree centigrade. I think it should be at 4 degree
> centigrade. Is this correct?

Check the manual for the TA cloning kit. I don't think it works with 
phosphorylated inserts. At least that's what I seem to remember with the 
TOPO-TA products.