Team:Shenzhen BGIC ATCG/notes

Timeline

Attributions

Work Design:

Jianhui Gong as a team leader and K2 as our instructor draft our project "Cell Magic".

Experiments Conduct:

Li Xiang Li,Xu Yanhui,Wu Fanzi, Yu yang from SCU: responsible for targeting peptide,XFP,terminators design and experiments.

Chen Shihong,Gu Chenguang, Lu Yanping, Liang Jiale,from South China University of Technology, are responsible for the alternative splicing Src1 and Mer2 intron design and experiments.

Zhu Shuang, Lin Kequan from Wuhan University and Wei Wei, Zheng Bingwei, Yi Lan in HUST work together for the promotors.

Guan Rui from SEU and He funan, Wang Rui, Lin Li,Zhang Yaolei from UESTC made their efforts to the degradation parts.

Zhou Wanling,Zhang Aiping, Li Dongdong, the undergraduates in AHMU, joined the part ***

Chen Yichun of SCNU, Zhong Na of JNU work for the microfulidic part.

The SCNU student: Chen Chengxuan, Lin Qiongfen, Xie Qiaolin, worked for the cell cycle regulator Sic1

Modeling:

Liu Shuang Liu from SEU, Zhou Yang from SCUT, Jinchun Zhang from SCU, Qiu Bitao from BGI

Wiki Construction:

Zhang Jinchun and Zhou Yang

Protocols

Protocol of chip-based XFP degradation rate detection in E.coli.

First of all, E.coli will be measured after shaking to about OD2.0（600）in the chip which was washed by plasma water, vacuum pumping. The bacteria liquid was pushed into the chip, letting the cells enter the small triangle. Then use the constant flow pump culture medium into chip (the laboratory constant temperature, can not guarantee the training environment, 37°E. coli slower growth). The medium speed is about 200ul/h. Finally we test the data after yeast fulled in the triangles.Using the IPTG medium, the new RFP expression was stopped and we can regard the lights as degradation tags' efficiency.

Protocol of Enzyme - labelled meter detecting the fluorescent protein intensity

First of all, take a certain amount of bacteria liquid, recovery to around OD0.6(600), ensuring the bacteria in the logarithmic growth phase. Diluted and then the bacteria transferred to 96 well plates, measured their fluorescence intensity. Red fluorescent protein using an excitation wavelength of 584nm, and its emission wavelength is 607nm.When measuring, we first detect the OD600 of each strain, removing the factor of bacteria number difference. Thus the fluorescence intensity cannot be altered by bacteria quantity. Then the measurement mode switching for measurement of fluorescence, fluorescence intensity.

References

Acknowledgements

== '''At the begining ''' ==

The BGI-ATCG iGEM 2013 team is a huge and diversity group consists of undergraduate from ten universities. Our team members’ major is really different, such as biotechnology bioinformatics, physical and even mathematics. Our work is partly assigned into each group made up by the students from the same university, which you can find in the group part in this page. And you can also browse about who and how everyone contributed to the project in this page. We really appreciate all our advisors and instructors that have assisted us throughout this project, without whom the project could not been carried out. We would also like to thank all everyone else who has helped us to achieve our project through put up advice or providing DNA, seeds, or other materials. Their contributions have helped us enormously. For a full list of acknowledgments, please see the bottom of this page.

=== '''Financial Support ''' ===

BGI College provides the totally funding including our team registration fee and competition travel fee.

SUSTC Biology Department and the BGI Unit of Synthetic Biology help us purchasing some materials.

=== '''General Support'''===

Cho-Kiu Wong at SMC, BGI Tech Solutions help us much to send and receive the Biobrick

Unit of Synthetic Biology at BGI trains team members about the basic experiment technology and knowledge which was really necessary for us.

=== ''' Material Support''' ===

Boeke Lab of John Hopkins Medical Institutions (Boeke Lab @ John Hopkins Medical) provide us pRS413, pRS414, pRS415, pRS416

Dr. Chi-Ming Wong (Dr. Chi-Ming Wong @ HKU) at Hongkong University provides us the plasmid yplac195 yplac181

Kai Tian and Yong Li at BGI, helping us with the CRIPSR system.

China_WHU has provided us the E. coli version of CRISPRi system.

=== '''At last''' ===

We really appreciate 1)the BGIC_0101 team for their cooperation, 2)the SCSTC help us for borrowing device and booking materials, and 3) SYSU, SUSTC borrowing us some experiments equipment.

=== ''' Lab Support''' ===

Unit of Synthetic Biology at BGI supports us with the lab for the most important cell and molecular biology experiments.

Dr. Ming Ni, team leader in BGI cancer group, let us use his lab for the microfluidic experiments.

Pr. Lingling Shui at south china normal university provides the microfluidic lab and materials for our experiments.

=== Cooperation college or university ===

Huazhong University of Science and Technology

Wuhan University

China University of Geosciences

South China University of Technology

South China Normal University

Jinan University

Sichuan University

University of Electronic Science and Technology of China

Southeast University

Qingdao University

University of Chinese Academy of Sciences