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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!!


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


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!!!


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