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End of the project
What happened to the Fungal track 'n trace project?
After several months of running with our two separate projects we noticed that our divided attention was taxing our ability to succesfully complete two projects at once. There were several problems with the creation of protoplasts required for the transformations. At the same time we were struggling with the creation of the knockout. We used a method which involved extensive use of PCRs. It took us several weeks instead of several days to get our PCR working, which delayed the creation of our knockout at a crucial moment. However, all these problems combined were not enough to convince our team to stop working on the fungal project.
For our ambitious project we decided to implement a synthetic construct in A. nidulans. The decision was made to synthesize this construct at a foreign company, in the hope of saving ourselves time. The constructs were made in silico and after certain small corrections the sequences were submitted to the company. The constructs were submitted on time, but soon enough the DNA synthesis company let us know they were struggling with putting the final construct together. We received our first part in time and succesfully transformed A. nidulans and confirmed that our first construct worked.
However, our synthesis company had problems with putting the rest of our constructs together. Apparently one of the synthetic sequences was toxic for the E. coli host. When verifying the sequence no faultless sequence could be verified. Huge deletions were found in the sequence, indicating that there was something fundamentally wrong with our sequence. Upon closer inspection we found four small separate repeats in our sequence, however we strongly doubt this is the root of our problems.
Due to these combined problems our DNA synthesis company struggled with delivering the synthetic constructs on time. Halfway the summer with no delivery of the parts expected within a few weeks and with several transformations being delayed we realized it was impossible to deliver anything before the wiki freeze. Despite the best efforts of our synthesis company, the decision was made to wrap up the fungal track 'n trace project. Suzette continued to work on several microscopal aspects of fungal hyphae, but basically all of our man and womanpower shifted to the Synchroscillator project where we managed to get beautiful results.
