HUMAN PRACTICE

SCU-China Header

COLLABORATIONS

Peking

SCU-China needs a signal to represent our protein, while Peking needs indigo for their application. We have given each other the useful plasmid. SCU-China got the yeGFP, which can express the protein optimized for yeast. And Peking got the pET21-FMO. FMO can express a key enzyme in synthesis indigo, it was used in CRISProgrammer, our project last year.

SYSU-Medicine

We cooperated with the team of SYSU. For they needed the samples of Southwest China, we helped them to spread their questionnaires. And they assisted in promoting the video clips made by ourselves. Both the two teams benefited from the collaboration.

Tianjin

As we all know, the iGEM team of Tianjin University, as a senior yeast team, has used yeast as a chassis for many years and has rich experience in yeast transformation and culture. During the CCiC in 2019, we exchanged information on the electroporation and plasmid extraction of Saccharomyces cerevisiae with Tianjin University. The members of Tianjin University provided us with some suggestions as shown:

1. When performing yeast electroporation, the gradient of voltage and conversion time can be set, and parallel experiments can be set to reduce the error and explore the most suitable electroporation conditions.
2. Some protocols for yeast electroporation may give a mixture of 50 μl of yeast fluid and DNA in an electric rotor, which sometimes causes an electric spark when electric shock. It’s a good idea to raise the mixture fluid to 100 μl.
3. When culturing the transformed yeast, it is not necessary to culture to OD600=1.2. Plasmids can be extracted after a 12 hours YPD liquid medium culture.
4. In the yeast plasmid extraction, after adding Buffer YP II and YP III, remember not to shake the tube, one or two slight turns is enough to mix the liquid. Vortex may cause the yeast genome to be interrupted, affecting the results.
5. If the colony PCR of yeast has no result for many times, it may be considered to put the yeast fluid in the boiled water bath and ice bath several times to improve the colony PCR.

Based on the recommendations of the Tianjin team, we have revised our experimental procedures and the results have indeed improved to some extent. We would like to thank the Tianjin team for their suggestions and wish them excellent results in the iGEM 2019.

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