If+we+change+the+restriction+sites+of+the+vector+or+insert,+how+will+we+do+this?

Andrey and I discussed 3 possible strategies.

1. Cut from outside of the desired restriction sites. Then do several In-fusion reactions which would also generate a stuffer. The In-fusion reaction would have a total of 4 pieces (vector backbone, outside restriction site to inside fragment, stuffer piece, inside restriction site to outside fragment) 2. Cut with normal restriction sites and then clone stuffer fragment in with in-fusion. 3. Cut with normal restriction sites in default plasmid, and then perform an in-fusion to clone scFv genes in which would result in another amplification of the library.

Here's a scratch paper we used while discussing some of these ideas.