Our team's goal in the lab was to complete site-directed mutagenesis on the pSB1C3-asPink plasmid to elucidate essential amino acids for the colour.

Mutagenic primers were designed to change T94 and T95 in asPink to Q94 and I95. A Hpy188III site was removed using a silent mutation to change the restriction pattern of the mutant plamsid.
fwd primer
5' - GTACCCAGATTTATGAAGATGGCGGTTTTCTGACG - 3'
rev primer
5' - CATAAATCTGGGTACGTTCCCAGGTAAAGCC - 3'

1. Mutagenic primers are used during PCR to introduce point mutations to the new plasmid.
2. The PCR reaction is treated with DpnI to digest the orignal non-mutant plasmid.
3. The PCR product is transformed into E. coli and grown on selective media.

All protocols can be found here.