|
|
|
Attributions
|
|
From planning, researching and programming to fundraising, human practices and, of course, wetlab work, the iGEM competition is a huge (and rewarding) challenge that pushes students to hone their skills and learn new ones. As we are a team with a diverse background, we divided tasks to best match our member's experience and interests. Sometimes, that meant doing what we do best; other times, it meant learning something new. When difficulties arose, we turn to a supervisors for help and guidance. Here, we would like to creit our team member's for the specific roles they playing into making CascAID and to thank our supervisors and external help for their important contributions to our project.
|
Molecular cloning
Due to our background, most of us had experience in molecular cloning and after patiently explaining our physicist Katzi what a plasmid was (among other things) we were capable of doing all the cloning ourselves. Almost every member of our team helped with the cloning in some way, but most of the work for the Cas13a expression strains of E. coli came from the hands of Ludwig and Christoph. The cloning of our biobricks was done by Rob and Max, who also cloned the constructs needed in the Intein-Extein read-out. Florian and Christoph teamed up to make the parts necessary for the AeBlue read-out. Gene and primers design was done by those responsible for the cloning and proofed by our supervisor Aurore
|
Protein purification
Our three Cas13a proteins -Lsh, Lwa and Lbu- were purified thanks to the hard work of Ludwig and Max, who along with Aurore's feedback designed and cloned the parts necessary for the addition of the TEV-target-sequence. The purification was performed by them two together with the help of Milica and Benedikt
|
Targets
The original idea of using 16s rRNA from E. coli as a target for our initial experiments, along with many other things, came from our supervisor Aurore. For the other targets that cover a wider spectrum of pathogens, Dawafuti was the main responsible for choosing and designing them when needed. The target sequences from viral origin were a kindly given by Killian Vogele, a PhD Student from Prof. Simmels group. Dawafuti and Julian experimented with the diverse RNA extraction and purification methods for detection with Cas13a.
|
Interlab
Erika and Dawafuti took the interlab studies in their hands and, after being introduced by our supervisor Lukas to the machines, they two performed all experiments.
|
|
|
|
|
