INTERLAB STUDY 2018

results of our participation in the interlab study

Introduction

Being the biggest interlaboratory study done in synthetic biology, iGEM's InterLab Study aims to develope a robust GFP measurement procedure.

The 5th InterLab Study happening this year wants to establish an universal GFP measurement protocol. Following different fluorescence protocols from labs around the world will help us understand how reliable different plate readers are, how different people work and how close these worldwide results are one to another.

Materials

Methods

Following the iGEM_HQ Protocols and with the aid of the provided materials from the iGEM Distribution Kit and from our laboratory, we were able to perform the calibration ^[2] and cell experiments ^[3]. The same pipettes, plates and volumes were used to avoid calibration errors.

Results

Calibrations

Figure 1: Particle standard curve after measuring the Absorbance (OD600) from different LUDOX CL-X Solutions

Figure 2: Fluorescence standard curve after different Fluorescein concentrations.

Cell experiments

After the realisation of the calibrations experiments the cells were transformed with the required devices and LB Plates with CAP (see Table 1). All the transformations with the provided plasmids ^[1] except with device 5 succeeded at the first try. That made the whole process really fluent and easy. After a second successful transformation with the device 5 we could start the measurements.

Figure 3: Plates after the transformation of the *E. coli* with the desired device.

Figure 4: Graphic representation of the absorbances from different devices, controls and blank.

Figure 5: Graphic representation of the fluorescence emition from different devices, controls and blank.

Conclusion and experience with the InterLab Study 2018

All devices showed after the measurements positive growth and fluorescence. Devices 3 and 5 showed the poorest fluorescence. Their results were similar to the LB + Chloramphenicol and the negative control. The transformation of device 5 already presented problems but after that the OD600 was really similar to the other devices. Device 4 showed the best fluorescence results, but with a less OD600. The fluorescence from device 1, 2 and 4 were even better than the positive control.
Being part of the the InterLab study was a great and enriching experience. All the materials provided by the iGEM_HQ were simple to understand and use. More than ever collaborations between scientists and joining forces are the most important thing to enable more development in scientific research. With Interlab Study could we learn this values and hopefully as a group of young scientists we will apply them in our future work.

Team:Aachen/InterLab