The rules for designing primers are similar for both Gibson Assembly and NEBuilder HiFi DNA Assembly. Any primer designed for Gibson assembly will work with NEBuilder HiFi. However, some primers that work with NEBuilder HiFi may not work with Gibson because unlike the Gibson Assembly Master Mix, NEBuilder HiFi DNA Assembly Master Mix can remove 3' end mismatches. Tools which design "Gibson" primers may not allow the design of primers that take advantage of this ability of NEBuilder HiFi. Though it it is possible to use other tools, NEB recommends using our free tool, NEBuilder Assembly Tool, because it can design primers that consider the vector ends generated by a restriction enzyme digestion.