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PCR and topo problem - (Jan/26/2006 )

I want to insert my fragment into Topo vector, but first I want to insert a linker at the ends of PCR products. So I do my PCR with oligo carrying the linkers and then I do a Topo assay. After transformation I can detect my pcr product in Topo colonies, but when i sequence my pcr product the linkers have disapear!! huh.gif
somebody can explain me what happenned????

-Fredo-

Is this extra sequence that you have added to the 5' end of your primers? This should be there!

-tap14-

Yes I know , there should be there , I really dont't understand what happened... the mysteries of science...

-Fredo-

Have you sequenced your PCR product, to make 100% sure your primer has the linker?

-vairus-