There are three collaborations our team with other teams.
Collaborate with the team TUST_China
What help did we give them?
TUST_China shared their improved Tetracycline HPLC Protocol; They hope that we can help them take water samples and measure the concentration of tetracycline in water; further help TUST_China to improve the mapping of tetracycline pollution levels in waters in China.
After a few days of measurement, we obtained the results of tetracycline in the waters near the Beijing University of Chemical Technology and sent it to TUST_China. It can be seen that tetracycline pollution in Beijing still exists.
What did they help us?
Due to the lack of experience in measuring fluorescence values in our laboratory, we can't correctly grasp the time of fluorescence measurement, and the fluorescence plate is not enough. Our laboratory microplate reader is old and cannot guarantee accuracy, so in order to save time and less Taking a detour, we designed a quantitative experiment and then sent it to TUST_China to commission them to help us measure the fluorescence values of different fatty acid gradients. We designed four control groups. The first group only added 50 ul of 20% arabinose solution as a control; the second group added 50 ul of 20% arabinose solution, and after 1 hour, 100 ul of different concentrations of fat solution were added, and the fatty acid concentrations were 25 mM, 50 mM, 100 mM, 200 mM, 400 mM, respectively. The third group only added fatty acid solutions of different gradients, the concentration was the same as that of the second group, and the fourth group was the blank control group.
TUST_China helped us to analyze the fluorescence values of the bacterial culture after eight hours of cultivation as follows:
After receiving the fluorescence value measured by TUST_China, we can analyze the data of the group. The fluorescence value of the second group is higher than that of the first group of arabinose-only control group and the fourth group of blank control group. It indicates that the fatty acid operon plays a role, and the third group only adds fatty acids to produce fluorescence values, which may cause fluorescence leakage.
We are very grateful to TUST_China for helping us complete the experiment of measuring fluorescence, and have obtained good experimental data, which has advanced the progress of our experiments. At the same time, in addition to the experiment, the two teams often exchanged ideas on WIKI and other aspects to promote common progress. It is a great pleasure to work with TUST_China and hope that the friendship between the two teams will last for a long time.
Detailed information about the team-TUST can be found in the here: TUST_China.
Collaborate with the team UESTC-China
What help did we give them?
After introducing the gene plasmid containing GroES and GroEL into DH5α strain, UESTC-China verified that the growth of the recombinant strain was significantly better than that of the original strain at 1% butanol concentration. However, the single experiment was not convincing, so UESTC-China invited us to repeat the experiment to ensure the repeatability of the experiment.
We helped UESTC-China measure the effect of the expression of the GroESL gene on butanol tolerance in the DH5α strain. The effect of positive and negative samples of UESTC-China on butanol tolerance was measured. (The positive sample is E. coli DH5α with the GroESL gene introduced, and the negative sample is the DH5α wild type strain).
The measured data of the Abs600 within 24 hours are as follows:
What did they help us?
One of the keys to our experiment was the ability to construct a plasmid of interest. The operon and the vector plasmid we constructed are the key to this step. The ends of the operon are BglII and XhoI digested ends, respectively, which are complementary to the vector plasmid.
Thereafter, we performed an enzymatic ligation of the operon with the vector plasmid fragment under the conditions of a 16 degree reaction for 5 to 6 hours. Thereafter, we transformed the enzyme-ligated plasmid into E. coli top10 strain and observed its growth (the AMP resistance gene was carried on the vector plasmid to confer selectivity). The expected result is 1, 3, 4 groups of long bacteria, and 2 groups are not long bacteria. However, we conducted a number of tests and all ended in failure.
So, we contacted the cooperation team TEAM UESTC-CHINA to discuss the reasons for the failure and the solution. Finally, TEAM UESTC-CHINA proposed changing the enzyme ligation conditions, changing the enzyme ligation conditions to a 4 degree reaction for 22 hours (overnight reaction) for enzyme ligation, and proceeded to verify the work.
TEAM UESTC-China verified the results of the enzyme connection
In the end, we used the solution provided by TEAM UESTC-CHINA, and the experiment was carried out. The experimental results are very good. After that, we were able to carry out the subsequent operations.
I am very grateful to TEAM UESTC-CHINA for helping us, so that we have taken a lot of detours.
Detailed information about the team-UESTC can be found in the here: UESTC-China.
Collaborate with the team Tsinghua
Our team visited the IGEM team at Tsinghua University. Everyone exchanged ideas and spent an unforgettable morning together.
Visit the IGEM laboratory
We visited the IGEM laboratory of Tsinghua University. We learned about some experimental sites and equipment at Tsinghua University Laboratory.
Project exchange party
We exchanged learning in the conference room with members of the IGEM team at Tsinghua University.