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

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