Team:CIEI-BJ/InterLab
In order to meet the requirements of the Bronze Medal for iGEM, our team completed the InterLab Measurement Study this July, which aims to improve the measurement tools available to both the iGEM community and the synthetic biology community as a whole. In accordance with iGEM 2018 InterLab Study Protocol, we finished the whole process using plate readers in the labs of Beijing Science & Technology University, and finally achieved desirable results.
Our team members Zhenyu Xiong, Haolun Cheng and Zheyu Guo participated in and completed this experiment. The process and results of the experiment is as set out below.
Plate reader（multifunctional enzyme marker）
Devices：
| Positive control | BBa_I20270 |
| Negative control | BBa_R0040 |
| Device 1 | BBa_J364000 |
| Device 2 | BBa_J364001 |
| Device 3 | BBa_J364002 |
| Device 4 | BBa_J364007 |
| Device 5 | BBa_J364008 |
| Device 6 | BBa_J364009 |
(Provided by the committee)
96 well plate, black with clear flat bottom
Fluorescein (provided in kit)
LUDOX CL-X (provided in kit)
Silica beads - Microsphere suspension(provided in kit)
Some conventional experimental appliance
Interlab's protocols are detailed and easy to manipulate, but the last step dilution concentration of the bacterial solution is too low, and the lowest level of concentration in the medium is almost non-colony. We repeated the experiment several times before we got the result we expected.
Since there are variations among different measuring instruments, we found out the standard value in our own apparatus by using three sets of unit calibration measurements: an OD600 reference point, a particle standard curve, and a fluorescein standard curve. The results we achieved are shown as below.
Data
Data
Data
For cell measurement, we transform eight plasmid to agar plat and then did cultivate the culture overnight. Then follow the protocol, we did series dilution and then measure the Abs 600 and fluorescence. After 6h, we measured the Abs600 and fluorescence again .
Data
During the process of InterLab, we were introduced to the multifunctional enzyme marker for the first time and learned how to use it, which greatly improved our experiment skills. We failed in the first two experiments due to our lack of experience but we finally achieved success in our third try, and we are really proud of it!
