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Can I PCR amplify a ligation product directly - (Apr/06/2005 )

who have ever done this before, any suggested protocol for it, thanks!!

-shy-

I haven't done the exact experiment as you described (you want to amplify two ligated fragments right?).
But I did PCR to check ligation of plasmid before It works. I just take 1 ul ligation mixture as template and run the PCR. I noticed that sometimes beside the product band, I also got smear.

good luck

-bullfrog-

hi, bullfrog, thanks for your anwser, I have already done the PCR, it only gives a smear, but no band. since my ligation product contain a promoter library, I need to amplify the ligation product to ensure that as many promoter as possible are contained since transformation can only give single promoter clones.

any suggestion are welcome, thanks!!!

-shy-

hi, shy.....I don't understand what do you mean. Could you explain again. Is overlapping PCR or nested PCR?

Regard,
Ali

-Ali-