Team:TU Eindhoven/Experiments
Experiments
Here you can find and download all the protocols that were used for our labwork.
General Protocols
Preparation of general necessities 🔗 Preparation of antibiotics, growth media, stock solutions and more. NanoDrop 🔗 NanoDrop is used to determine the concentration of DNA – and protein samples. Agarose Gel Electrophoresis 🔗 Agarose Gel Electrophoresis is used to determine the length of DNA samples. SDS-PAGE analysis 🔗 SDS-PAGE is used to determine the size of a protein. Gel extraction 🔗 Gel extraction is the silica-membrane-based purification of DNA fragments from gels.
Cloning, Mutagenesis & Gibson Assembly
PCR Amplification 🔗 PCR Amplification is used to amplify a DNA sequence. PCR Purification 🔗 PCR Purification is the purification of a product from a PCR reaction. Digestion 🔗 Digestion is the cleaving of a vector and insert using restriction enzymes to prepare them for ligation. Ligation 🔗 Ligation is the process of ligating an insert into a vector. Vector Linearization 🔗 Vector Linearization is the process of linearizing a vector, after which it can be used for Gibson Assembly. Gibson Assembly 🔗 Gibson Assembly is the ligation of gBlocks into a linearized vector.
Transformation, Protein Expression & Purification
Transformation into competent cells & plating on agar plates 🔗 Transformation is the introduction of a desired plasmid into bacteria to amplify it. Colony picking & colony PCR 🔗 Colony picking & colony PCR are used to determine the present of insert DNA in plasmid constructs. Sequencing at BaseClear 🔗 Sequencing is used to determine the sequence of the amplified and transformed DNA. Small Culturing 🔗 Small culturing is used to amplify a specific bacterial colony after transformation. Protein Expression 🔗 Protein Expression is the induction of protein production in bacteria from small culture. Protein Purification 🔗 Protein Purification is the purification of the desired protein using purification tags.
Testing
Bioluminescence Measurements 🔗 Bioluminescence Measurements are used to determine the bioluminescent activity of a desired compound.
Phage experiments
Preparation of media 🔗 Preparation of M9-based minimal medium for bacterial growth used in phage experiments. Bacteria multiplication rate determination 🔗 Bacteria multiplication rate determination is important for the determination of certain parameters used in the model. Determination of adsorption rate 🔗 Determination of the adsorption rate, which will be used in the model. Infection Assay 🔗 The Infection Assay is used to determine several parameters for the model. Phage Titer Determination 🔗 Determination of the phage titer of a certain solution.
