Team:HZAU-China/Labnotes

Parts connection and standardization:

Author: Zheng Yihui

Members: Zheng Yihui, Zhang Yajing, Liu Zhixiang, Luo Xudong, Guo Xiaoxue, Wang Lu, Li Siqi.

Introduction:

We have eight gene circuits to construct (as shown in the figure.1), and after finishing these circuits, we will integrate some of them into one plasmid to form two repressilators and two detection devices (as shown in the figure.2a and figure.2b).

May

Content:

we found the parts we needed in the Plate Kits and did a lot of transformations to get the plasmid for subsequent experiments.

Our project includes three systems---input system, processing system and output system, which are constituted by eight gene circuits, and each circuit is composed of at least three parts(as shown in the design overview of our project). We take almost four months to construct all of them by the standard method of biobrick connection and test them whether they work as expected or not by detecting fluorescence intensity, and definitely, it is totally a long and tough road. During the process, we conduct all kinds of experiments like double digestion, ligation, transformation and even some chemical experiment to put one gene in front of or behind another gene and synthetize the material we need. Besides, we make our greatest efforts working on testing our parts and analyzing the data we get. Unexpectedly, we find a phenomena related to the unexpected recombination of lox71 in DH5α and of course, we have done some testing to verify our assumption. Anyway, the road to success is laborious, but we still stick to our goal and finally make it by ourselves. The following is the methods of our construction and the record of our experiments (recorded by different writers).