Team:HUBU-Wuhan/InterLab
The HUBU-Wuhan iGEM 2018 team is participating in the Fifth International InterLaboratory Measurement Study in synthetic biology.
Reliable and repeatable measurement is a key component to synthetic biology. One of the big challenges in synthetic biology measurement has been that fluorescence data usually cannot be compared because it has been reported in different units or because different groups process data in different ways. In the previous studies, the measurement committee showed that by measuring GFP expression in absolute fluorescence units calibrated against a known concentration of fluorescent molecule, we can greatly reduce the variability in measurements between labs. This year, we are invited to answering the following question: Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD?
Strain
Plasmids
Meterials
Methods
OD600 Reference Point
Particle standard curve
Fluorescein standard curve
More cell measurement data see InterLab_2018_HUBU-Wuhan (download)InterLab
Background
Materials& Methods
Escherichia coli strain DH5α
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
-1ml LUDOX CL-X
-150 μL Silica Bead (microsphere suspension)
-Fluorescein
-1x PBS (phosphate buffered saline, pH 7.4 - 7.6)
-ddH2O
-LB (Luria Bertani) media
-Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH)
-50 ml Falcon tube
-1.5 ml eppendorf tubes
-Ice bucket with ice
-Micropipettes
-Micropipette tips
-96 well plates, black with clear flat bottom preferred
→Calibration protocol (preview)
→Cell measurement protocol (preview)
Results
