- NCTU_Formosa 1st online meetup
- Culturing our Pinus armandii callus in NCTU_Formosa's bio lab
- NCTU_Formosa 2nd online meetup
- Helping NCTU_Formosa on the measurement of O.D value
NCTU_Formosa 1st online meetup
This meet-up was for discussing forming a partnership. Besides exchanging ideas about the project, we asked them if they could help us with conducting qPCR of nematodes and lending us the plant growth chamber. And we would help them with collecting and analyzing the saliva of our school's stray dogs to provide the environment to test their product—Denteeth (dental bone for pets).
We accomplished the part of plant callus and culturing a species of bacteria Komagataeibacter xylinus. This bacteria can produce konjac, NCTU_Formosa planned to use konjac for delivering their product. Because of the difficulty of analyzing each component precisely in saliva, we replace the collaboration into culturing K. xylinus for them. (Fig.1)
NCTU_Formosa → NTHU_Taiwan
NCTU_Formosa help us on culturing the callus of Pinus armandii
We need the callus of P. armandii to test if our target gene was successfully transferred into plant cells and if it worked. We got all the experimental material and seeds, except for the plant growth chamber. Because the two schools are adjacent, they generously lent us the plant growth chamber from their lab and allowed us to conduct experiments there. Their teachers and members had plentiful experiments on culturing plant callus, they also gave us some advice about how to culture plant callus. The following figures show the course of events. (Fig.2~4 )
We went there to passage our callus every week and successfully got the plant callus. Figure 5~7 are the growth records for the first week. In the first week, the callus grew slowly. From the second week, we can see the callus growing bigger (Fig.8). Eventually, we use the callus, which NCTU_Formosa cultivated for us in our experiments!
NCTU_Formosa 2nd online meetup
To do further discussion and confirm the progress of our partnership, we had the second online meetup(Fig.9). Because of the covid-19 outbreak, the Food Industry Research and Development Institute in Taiwan cannot deliver NCTU_Formosa's order of K. xylinus in time. Thus, we decided to change our collaboration of culturing K. xylinus to the measurement for model validation.
NCTU_Formosa wanted to confirm whether the model they designed conformed to the actual situation; they needed an extra set of data in the model to be more referential. Although, the limit of time and lack of facility cause difficulties in execution. On the other hand, we got the spectrophotometer in our lab and can execute the measurement every hour automatically, which saves much time, so we decided to help NCTU_Formosa measure whether the O.D value of the growth model fits the prediction of the model they designed.
NTHU_Taiwan → NCTU_Formosa
NCTU_Formosa wanted to verify whether their models for predicting the growth of E.coli and its expression conform to the actual situation. They need to measure the O.D value of E.coli and the relative fluorescence of RFP and GFP. To obtain intensive data to compare with the simulated growth curve, it needs to be measured an hour at a time in 24 hours. However, NCTU_Formosa had no machine that can regularly measure these three values. To solve this difficulty, we used machines in our laboratory to assist them in obtaining these values to complete the validation of their model. The following are photos of their members coming to our laboratory. (Fig.11~12)
We got the result after the measurement (Fig.13). As you can see below, the O.D. value of E. coli had not changed significantly. It showed that E. coli had reached a growth balance. And Moreover, we could find found both the expression of GFP and RFP values stayed in a range, indicating their product —DenTeeth could stably produce protein.
They ensured the stable expression of E. coli and have enough data to compare with the simulation of the model. They also adjusted the parameters to complete the model validation. With this modified model, they can predict the growth and expression of DenTeeth more accurately. And because of the partnership, we got a chance to learn more about their project!
Fig.13(a)～13(c) The result of measurement.