Team:Tuebingen/Labwork
Labwork
Everybody's a mad scientist, and life is their lab. We're all trying to experiment to find a way to live, to solve problems, to fend off madness and chaos.- David Cronenberg
Introduction
BoNT C - Liscense to enter
As stated, we modified botulinum toxin in a way that lead to its detoxification.
This, as well as the coupling of our detoxified BoNT C to other substances were the main aspects of our lab work, next to our special cell culture to test our library on neuron-like cells. The whole process we went through and the different methods we used to achieve our goals are described below.
An other important part of every iGEM year is the InterLab Study, which is one of the Bronze-Medal Criteria.
If you are intersted in our Bioinformatic part: BioInfo
Methods
Working neatly is very important to every scientist. Because of this, here we're describing our methods. You can click any of the tabs to see what we did in each laboratory.
Generating Electrocompetent Cells
Transformation of Cells
Mini Preparation of Plasmid DNA
Maxi Preparation of Plasmid DNA
Restriction Digest and Ligation
Expression of the protein and harvesting of the bacterial culture
His-tag protein purification
Strep-tag protein purification
Set up buffer
Desalting samples
SDS gel preservation
Western blot
SHSY5Y basic cell culture
SHSY5Y freezing and thawing
SHSY5y cell splitting
Differentiation protocol
Explanation of distillation
Explanation of chromatographie
Explanation how to dry solvents
Synthesis of the Thiol-Esli
Synthesis of the Disulfid-Esli
Synthesis of the Azid-Donor
Labbook
Here you can read our Labbook. It is the place, where we described everything that we did, and when. If you are interessted in our workflow, you can read everything below.
Everything done in the MolBio Lab
Labbook: protein purification, expression and toxassay
Labbook: Cell Culture
Labbook: Organic synthesis
Interlab
In this year, our team again decided to participate in the fifth international iGEM InterLab study, which aims to identify and correct the sources of systematic variability in synthetic biology measurements. The overall goal is that eventually, measurements taken in different labs will not underly the problems of variability due to different measurement environments or devices anymore, but will be reliable and comparable for all members of the science community.
The main question the InterLab Study 2018 tackeled was:
Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD?
If you are interested to read more about our InterLab study results, click here:
