Am working with cloning, and plasmid has an Sap1 restriction site at Forward and reverse (purchased the cut plasmid from an company, they call it Electracloning) and the insert gene has forward and reverse ends of Sap1. After cloning the gene into the plasmid, Sap1 restriction enzyme will not cut the plasmid in Tango1x buffer, my question is how to make Sap1 cut the circular plasmid, what changes must be made for the restriction enzyme Sap1 to work ?
Thanks in advance.