We participated in the InterLab study organised by CCiC and Genescript. We characterized the performance of previous parts, TCI42, BBa_K2572001, and TlpA39, BBa_K2572000. We also characterized the performance of novel parts, TEVts6#, BBa_K2969001, TVMV protease, BBa_K2969006, and PheOH, BBa_K2969025.
We participated in the "Clone It" InterLab study, which was organised by CCiC and Genescript, to test the Gibson assembly/In-fusion HD assembly kit and provide feedback.
Interlab is a large-scale collaboration through which to provide more available tools to both the iGEM community and the synthetic biology community as a whole. This year's collaboration was initiated by CCIC，aiming at testing the effects of a new kit. The main test was a new DNA ligase. As a participant, we did the following experiment and finally reported back to the sponsors the data we obtained.
Complete step for plasmid construction
We selected the construction of a plasmid required for the project to carry out the experiment. Starting from the initial PCR process, we performed a complete set of plasmid construction in strict accordance with the provided experimental procedures.
New DNA ligase use and effect detection
Since the biggest difference of the kit is the enzyme provided for DNA ligation, in the DNA ligation step, we use the novel reagents provided in the kit according to the usage, and then carry out the subsequent plasmid construction operation, and Finally, it was verified by colony PCR that the enzyme was effective and got a good feedback data.
CHARACTERIZATION OF PREVIOUS PARTS
We characterized the performance of TCI42, BBa_K2572001, and TlpA39, BBa_K2572000, by testing their temperature responding curve using flow cytometer this year. See more information from BBa_K2572001and BBa_K2572000.
We inserted different number of TAG codon to sfGFP at different site to see the effect of TAG codon to the expression of sfGFP. The result shows that TAG codon can inhibit the transcription of sfGFP effectively. See more information from BBa_I746916.
CHARACTERIZATION OF NOVEL PARTS
About TEVts#6, we measured its temperature response curve. About TVMV protease, we measured its cleavage efficiency to CI434-TVMVsite induced by a series of concentrations of IPTG. Finally, about PheOH, we conducted kinetic analysis and got its kinetic parameters.