Team:SCU-WestChina/InterLab

index

Interlab
How close can the numbers be when fluorescence is measured all around the world?
Introduction

Over the past three years, iGEM has advanced the frontiers of science with the biggest interlaboratory studies ever done in synthetic biology. The SCU-WestChina iGEM team is honored to participate in the Interlab study this year.

The aim of the Interlab study is to analyze the measurement data of fluorescence with a detailed protocol, but different lab equipment and environment. In this way, researchers are able to get an access to compare fluorescence data from laboratories all over the world, in a standardized way.

Protocol

InterLab 2017 Plate Reader Protocol was followed. The device utilized in our Interlab experiments was BioTek Cytation™ 3.

For this study, following constructs are used:

•Negative Control: I20270

•Positive Control: R0040

•Test Device 1: J23101.BCD2.E0040.B0015

•Test Device 2: J23106.BCD2.E0040.B0015

•Test Device 3: J23117.BCD2.E0040.B0015

•Test Device 4: J23101+I13504

•Test Device 5: J23106+I13504

•Test Device 6: J23117+I13504

All devices and parts were obtained from the iGEM 2017 InterLab Distribution Kit. Devices were made with a pSB1C3 plasmid backbone and transformed into Escherichia coli DH5α.

Data & Result
Fig. 1 OD600 curve for each replicate, which suggests that most devices had no influence on the replication ability of the cells, while Device 4 and Device 1 showed much lower growth curves.
Fig. 2 GFP fluorescence data of each device and replicate measured with an excitation of 485 nm and emission of 530 nm
Fig.3 The ratio of fluorescence to growth for each Interlab device.

Results from the Plate Reader indicated that the promoter of the Device 4 is strongest, followed by the promoter of the Device 2 and Device 1.

The lowest Intensity was detected in Device 3, Device 5 and Device 6.