Team:Stanford/InterLab
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Reliability and reproducibility are critical for successfully executing synthetic biology. Nevertheless, differences in measurements between labs persist. The Fifth International InterLab Study continues the work of prior InterLab studies and seeks to establish an effective and reproducible measurement procedure for GFP, thereby reducing variability between labs. This year’s InterLab explores whether normalizing to colony forming units (CFUs) reduces variability between labs relative to normalizing to optical density.
We used chemically competent DH5-alpha E. coli cells ordered from New England Biolabs (NEB).
Plasmids:
Additional Materials:
Plate Reader Model: Synergy H1
Plate: 96-well plate, black with clear bottom
Optics: Bottom
Absorbance: 600nm
Read Speed: Normal
Read Speed: Normal
Excitation: 485
Emission: 528
Gain: 50
We followed the calibration protocols directly as outlined in the iGEM InterLab Protocol in order to get baseline measurements for our plate reader.
Calibration 1: OD600 Reference point - LUDOX Protocol
Calibration 2: Particle Standard Curve - Microsphere Protocol
Calibration 3: Fluorescence standard curve - Fluorescein Protocol
We followed the cell measurement protocol directly as outlined in the iGEM InterLab Protocol
We followed the cell measurement protocol directly as outlined in the iGEM InterLab Protocol
