Team:SCUT-China A/InterLab




        All of the 2017 iGEM teams are invited and encouraged to participate in the Fourth International InterLaboratory Measurement Study in synthetic biology. Our team join in this study which aims to standardize the measurements of GFP’s fluorescence. And this year our task is measuring some RBS devices that are intended to make gene expression more precise and reliable. In this study, we were using plate reader for measuring.

Materials and methods

All the plasmid located in Kit Plate 7
1) Positive Control (BBa_I20270): well 21B
2) Negative Control (BBa_R0040): well 21D
3) Test Device 1 (BBa_J364000): well 21F
4) Test Device 2 (BBa_J364001): well 21H
5) Test Device 3 (BBa_J364002): well 21J
6) Test Device 4 (BBa_J364003): well 21L
7) Test Device 5 (BBa_J364004): well 21N
8) Test Device 6 (BBa_J364005): well 21P

2. Stain

E.coil DH5α

3. Materials

1) 1ml LUDOX (provided in kit)
2) fluorescein (provided in kit)
3) 1xPBS (phosphate buffered saline)
4) LB (Luria Bertani) media
5) Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH)
6) Glass tubes for cell grown
7) 1.5 ml eppendorf tubes for sample storage
8) 96-well plate, black with flat bottom
9) 96-well plate, transparent with flat bottom
10) pipettes

4. Machines

Tecan Infinite M200 Pro

5. Methods

Calibration protocols
1) OD600 reference point
2) Fluorescein fluorescence standard curve
Cell measurement protocol

OD600 reference point

Fluorescein fluorescence standard curve

Cell measurement


        Although we’ve finished this work, we made some mistakes. We are sorry about that there is something wrong with our data, because we used the Tecan_m200_Pro plate reader and we chose Optimal as our gain setting.