Difference between revisions of "Team:Munich/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 cloning and experiments.

RNA extraction

One of our two inital ideas for RNA extraction, Julian and Pattrick did multiple experiments trying to use beads to bind RNA. For our another option, Jorge experimented with the FINA extraction method as presented in McFall et al., 2015 and tried to adapt it for RNA extraction.

Read-out

As we looked into colorimetric read-outs for CascAID, our supervisor Jeff helped us brainstorm and suggested two interesting options. One of them we dubbed AeBlue read-out and was explored by Christoph, who did part of the initial cloning, and Florian, who performed all main experiments. The other was the Intein-Extein read-out. The cloning and experimets for this read-out were done mainly by Max with help from Sven. Rob worked in the gold nanoparticles read-out with help from Killian Vogele, a PhD student in Prof. Simmels' group. Killian also kindly gave us some linker sequences necessary for this read-out.

Lyophilization

Lyophilization, both of Cas13a and other components, was done by Rob, who reached out to Lady's name for guidance and equipment. With her support, Rob was able to use the machines at the Lady's institute to make the lyophilization experiments.

Hardware

Our hardware team consisted of four students: Katzi, Matthias, Sven and Max. Our fluorescence detector "Lightbringer" was conceived, designed and built by Katzi and Matthias, while Sven and Max developed "Heatbringer". Along the way, our supervisors Lukas and Benjamin gave valuable input and feedback.

Hardware

Our hardware team consisted of four students: Katzi, Matthias, Sven and Max. Our fluorescence detector "Lightbringer" was conceived, designed and built by Katzi and Matthias, while Sven and Max developed "Heatbringer". Along the way, our supervisors Lukas and Benjamin gave valuable input and feedback.