Team:NAWI Graz/InterLab

Introduction:

We took part in the iGEM Interlab Study where we worked with 6 different RBS to establish a protocol for GFP measurement in a plate reader.

Materials:

Plate reader:

TECAN GENios Pro

Plate reader Plates:

96 well black plates with clear flat bottom (Greiner)

Cell culture shaker:

INFORS HT MULTITRON

Devices:

Table 1: Test Devices of the Kit Plate

Microorganism:

E. coli K-12 DH5-alpha

Calibration material:

LUDOX CL-X
Silica Beads - microsphere

Methods:

We followed the protocol provided by iGEM HQ.

Comments:

We got the E. coli K-12 DH5-alpha from the Vienna iGEM Team (BOKU-Vienna)

We got the Chloramphenicol (in powderform) for the Interlab Study from the Munich iGEM Team.

We also took part at the Flow cytometrie and uploaded all the data to the dropbox

Plate reader configuration:

Gain: 600
Number of flashes per well (absorbance): 10
Number of flashes per well (fluorescence): 45
Temperature: 25°C
Emission wavelength: 535nm

Bandpass width: 25nm
Excitation wavelength: 485nm
Fluorescence reading: top optics

Results:

Table 2: Callibration with LUDOX Solution

Figure 1: Particle standard curve of the callibration experiment. Measurement by 600nm

Figure 2: Particle standard curve (log scale) of the callibration experiment. Measurement by 600nm

Figure 3: Fluorescein standard curve of the callibration experiment

Figure 4: Fluorescein standard curve (log scale) of the callibration experiment

Table 3: Calculated Table by the iGEM Exel File

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