Transformation with recombinant plasmids:
The PCR fragments from the Top A primers and the cp primers were ligated in separate reaction mixtures with the pGEM-Teasy vector, and then E. coli strain XL-1 Blue was transformed by heat shock with each reaction mixture. These cells were plated on 2 X YT medium plus amp and tet and X-gal and IPTG spread on the surface. This allows for blue/white selection. The plate below is from the transformation with PCR fragment from the cp primers. White colonies for both the top A and cp protein recombinant plasmids were saved. These cultures (each white colony is a culture) were grown on liquid 2 XYT + amp and tet overnight and the plasmids extracted by the standard miniprep DNA procedure.
Note the blue and white colonies on this plate. Reaction mixture of the vector = coat protein PCR fragment used in transformation. White colonies (5) were circled (black ink pen) on bottom of plate and then bacterial cells were transferred from each colony to new plates for storage at 4 C. The plates with the recombinant plasmid for the top A PCR fragment looked similar.
[back] [recombinant plasmid gel]