Primer mismatch bother me! - when enzyme sites added at 5' ends, mismatch increases. (Jan/06/2008 )
Dear all,
Before enzyme sites added, I amplified the fragments from the plasmids, I could get the bright clear target fragment, but the second step PCR adding enzyme sites at 5' ends of the primers, the results:
I could got the target fragment band, which was brighter than the backgroud, but also with smeared product, increased the annealing T, the result was the same.
-genejackie-
Can we have an image, please?
-swanny-