1. You have a 2 kb plasmid, and you want to insert a gene thatgives ampicillin resistance that is 500 bp in size. You also wantto insert a gene for penicillin resistance that is 435 bp in sizeand your PCR product, which is 250 bp in size. What would the totalsize of the plasmid be following the insertions?
2. Why would a different column, one unlike the silica gel purification column used in the protocol, cause anunsuccessful transformation?
3. What would happen if you forgot to heat shock the E. coli cells during the transformation reaction?