Team:FJNU-China/InterLab
Interlab
Overview
Reliable and repeatable measurement is a key component to all engineering disciplines. However, the number of cells in the sample is a variability in measurements. The goal of the iGEM InterLab Study is to identify and correct the sources of systematic variability in synthetic biology measurements. This year, our team take part in the fifth InterLab study, using normalizing to absolute cell count or CFUs instead of OD to reduce variability of fluorescence measurements.
Materials
Strain usedEscherichia coli DH5α
Plasmid usedNegative control: BBa_R0040 Plate 7 Well 2D
Positive control: BBa_I20270 Plate 7 Well 2B
Test Device 1: BBa_J364000 Plate 7 Well 2F
Test Device 2: BBa_J364001 Plate 7 Well 2H
Test Device 3: BBa_J364002 Plate 7 Well 2J
Test Device 4: BBa_J364007 Plate 7 Well 2L
Test Device 5: BBa_J364008 Plate 7 Well 2N
Test Device 6: BBa_J364009 Plate 7 Well 2P
MachineMolecular Devices SpectraMax i3x
Methods
We followed this protocol to do the Interlab Study.When we completed three of the calibration measurements, performing the cell measurements. Used the same plates, volumes and settings that we used in calibration protocol. We transformed E.coli DH5α competent cells with the 8 plasmids and picked 2 colonies from each of plates into 5 mL LB medium + Chloramphenicol. After culturing the cells overnight at 37°C and 220 rpm, we used plate reader to measure the Abs600 and fluorescence of samples at 0, 6 hours.
Result
CalibrationCalibration 1: OD600 reference point
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Analyse
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