Posted 10 April 2009 - 12:20 AM
i am designing a synthetic gene and i need to have Sal1and EcoRV restriction site at the 5' and 3' terminals.can i just insert the restriciton site of the enzyme on both ends and then design the whole gene..or do i need any additional base pairs in addition to these restriction sites on 5' and 3' ends....
Posted 10 April 2009 - 03:57 AM
Posted 10 April 2009 - 10:05 PM
I would suggest switching to a different enzyme than SalI, which has a very poor reputation for cutting PCR fragments.
XhoI and SalI form compatible overhangs