Team:Nanjing-China/Collaborations

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We have formed online collaboration with NEFU since September. We informed each other about our progress and helped each other with experimental puzzles. Some of the problems we tackled together are listed as follows.

♝:We plasmid vector can't be cut.

♗:It may be because of the enzyme had losed potency :

Plasmid was contaminated with protein
The plasmid may be dissolved in a solution that inhibited the activity of the enzyme

♝:What should we do to link three parts efficiently?

♗:Use isocaudamer.

♝: What should we do when we we want to dye the sample by Comassie blue staining and WB.

♗:When you add the samlpe, you should pay attention to the amount of it. Measure the value of OD to get the concentration of the sample you need and then add the sample. When making up polyacrylamide agarose gel, wash and dry the holder. In that way, you can get the gel free of bubble.

Vist the wiki of NEFU for more information at https://2017.igem.org/Team:NEFU_China/Collaborations

We obtained the plasmids from OUC-China at the beginning of October. We used the LiAc/SS carrier DNA/PEG method to transform yeast strain EBY100. The kit we utilized in the transformation is the Frozen-EZ Yeast Transformation Ⅱ^TM from ZYMO RESEARCH. We used the YPD(G418+) plates and incubated them at 30℃ for 4 days to allow the growth of transformants. We picked several yeast colons and grew them in 10ml YPD(G418+) broth. Then we monitored the growth rate of the recombinant yeast cells to specify the expression of MINI system at a particular living stage. The fluorescence and Abs600 of the four strains were determined by a microplate reader as shown in the picture.

We gave the result to the team,OUC. They are glad to see their results was repeated in our lab environment and revealed an ideal result.

In return, we asked OUC-China to help us validate the function of our formaldehyde whole-cell sensor. We wanted to see whether our plasmids could work under other conditions. Their works are shown at https://2017.igem.org/Team:OUC-China/Collaborations

Address

Life Science Department
#163 Xianlin Blvd, Qixia District
Nanjing University
Nanjing, Jiangsu Province
P.R. of China
Zip: 210046

Email

nanjing_china@163.com

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