Team:Bilkent-UNAMBG/InterLab

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InterLab Measurement Study

Introduction

Bilkent UNAMBG 2017 Team participates this year’s Interlab Study as well as the last year. The Fourth International InterLaboratory Measurement Study is an optional acitivity for Bronze Medal.

GFP is a widely used marker with many advantages. It is easy to use, cheap and does not require any substrate. Although it is used in many labs around the world, the measurement of GFP shows inconsistencies. Avoiding such situations and improve comparability is the major subject of this year’s Intelrab study. After gathering many GFP data from teams, preparation of a detailed protocol is aimed in this study.

Materials and Methods

Composite parts and a measurement kit has been given to all teams. First, measurement kit was used to standardization. Generating standard curves, we were ready to perform GFP measurements. Composite parts are composed of 8 plasmids(Positive Control, Negative Control, Test Device 1, Test Device 2, Test Device 3, Test Device 4, Test Device 5, and Test Device 6). Their part numbers are listed in the table. Before measurement, the plasmids had to be transformed into E. coli K-12 DH5-alpha. The cells were grown in LB medium with chloramphenicol. The measurement protocol has been performed with a plate reader SpectraMax M5.

The protocol can be reached by using this link.

Figure 1: Test Devices 1/2

Figure 2: Test Devices 2/2

Results

Figure 3: Standard Curve of fluorescein at 485nm excitation
and 530 nm emission wavelength

Figure 4: Standard Curve of fluorescein in log scale at 485nm excitation
and 530 nm emission wavelength

Figure 5: Average Fluorescence over 6 hours

Figure 6: Average Absorbance over 6 hours

The protocol can be reached by using this link.