Team:TU Eindhoven/Protocols

Protocols

For this year's iGEM competition numerous protocols were devoloped to guide our experiments and keep our documentation neat and tidy. Because these protocols can also be useful to other projects, we decided to publish them on our wiki. You can find information and download links on this page down below.

Genetic Engineering Protocols

The following protocols are used in the Biolab during the modification of bacteria.

Antibody Labelling	FACS for sorting with DBCO-TAMRA	Protein Expression Curve
Antibody Titration with FACS	Overhang PCR	Site Directed Mutagenesis
Bacteria Cultering for Microfluidics	PCR Purification of DNA Fragments	Vector Ligation
Colony PCR	Plasmid Amplification	Vector Transformation
Casting and Running 15% PAGE Gel	Creating Circular RCA Template	Plasmid Gene Digestion	Plasmid Purification
FACS for sorting with DBCO-PEG(10kDa)	Preparative Steps
Protein Expression	Labelling antibodies with 5/6-TAMRA-NHS

Chemistry Protocols

The following protocols are used in the chemical synthesis processes.

SPAAC reaction monitoring with UV-Visible Spectroscopy

Labelling Amine-Modified DNA with DBCO-PEG4-NHS Ester

Microfluidics Protocols

The following protocols are used in the Microfabrication lab for the production and running of microfluidic devices.

Droplet Separation	Oil and Water Phase (Polyacrylamide Beads)
Droplet Device Testing	Photolithography
Oil and Water Phase (Encapsulation)	Soft Lithography