Characterisation of An Existing Part
As our project had a large focus on fluorescence, we decided to characterise a part from the existing iGEM registry that produces red fluorescent protein in order to determine whether we could use it in our project. We decided against using RFP, however here are the results and methods used in the characterisation.
- The J04450 gene from the registry was cloned into three different plasmids (pSB1K3, pSB3K3, and pSB4K4) each with a different copy number.
- Each plasmid was transformed into electrocompetent MG1655-Z1
- Three colonies of each plasmid were then inoculated in LB and grown overnight.
- In the morning, each inoculation was refreshed in M9 + casein amino acid (500x dilution), induced with different concentrations of IPTG (0uM, 250uM, and 500uM IPTG) and grown for another 6 hours.
- They were then transferred into a 96 well plate (200uL per well) and measured their fluorescence every 15 minutes.
The results for our characterisation can be found on the J04450 part page in the registry.