From a PTH1R plasmid stock (100 μg/mL) a 1000x dilution was prepared in DNAse free water. Competent E. coli DH5α bacteria (ThermoScientific, USA) were thawed on ice. The bacteria (50 μL) were mixed with the diluted PTH1R plasmid (5 μL) incubated on ice for 30 min. The mix was incubated at 42 °C for 20 s and returned to ice for 4 min. Pre-heated S.O.C. medium (950 μL) was added, and this was incubated shaking (230 rpm) at 37 °C for 1h. After incubation the transformed bacteria were plated on LB ampicillin (100 μg/mL) plates and incubated at 37 °C overnight. The colonies grown on the LB ampicillin plates were picked and transferred to 5 mL of LB Broth (+100 μg/mL ampicillin) and further incubated shaking (230 rpm) at 37 °C for approximately 24 h. After 24h, the bacterial suspension was centrifuged for 10 min at 6000 rpm. Bacterial pellets were resuspended using the Monarch plasmid Miniprep resuspension buffer. Purification of the plasmid DNA was achieved with the Monarch plasmid purification kit following the manufacturers recommendations. The purified plasmid DNA was recovered in the elution buffer and characterised for DNA concentration, purity, and plasmid sequence.