The objective of this year’s study was to reduce lab-to-lab variablity in fluorescence measurements. This was done using two methods: normalising the absorbance to a known concentration of microsphere beads, and counting the colony forming units.
I Microsphere Beads Standard Curve
II Fluorescein Standard Curve
The parts used were:
Negative control BBa_R0040
Positive control BBa_I20270
Test Device 1 BBa_J364000
Test Device 2 BBa_J364001
Test Device 3 BBa_J364002
Test Device 4 BBa_J364007
Test Device 5 BBa_J364008
Test Device 6 BBa_J364009
The study was conducted using the 6 parts listed above.
The parts were in the plasmid pSB1C3 and were transformed into E. coli DH5a cells. The colonies formed were screened for fluorescence with appropriate biological and technical replicates using a plate reader and a flow cytometer. The overnight cultures were used in counting the colony forming units.
The 0 and 6 hour readings for two colonies that were screened per test device are depicted below.