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????
Is this extra sequence that you have added to the 5' end of your primers? This should be there!
Yes I know , there should be there , I really dont't understand what happened... the mysteries of science...
Have you sequenced your PCR product, to make 100% sure your primer has the linker?