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!!
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-