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From 2013.igem.org

Labbook |	Protocols |	Materials |

The following page(s) gives an overview of the protocols and possible modifications that where used and proven working during the iGEM PET.erminators project 2012. The corresponding materials can be found in the materials section.

Contents 1 In vivo 1.1 Escherichia coli 1.2 Comamonas testosteroni 2 In vitro 2.1 Nucleic acids 2.2 DNA 2.3 Protein

In vivo

Escherichia coli

• Bacterial cell culture
• Bacterial transformation aka TRAFO
• Cell counting/plating
• Chemically competent cells
• Colony PCR
• Electrocompetent cells
• Electroporation
• Glycerine Stocks
• Heat Shock Transformation
• Miniprep / Fermentas kit
• Miniprep / Promega kit
• Miniprep / Qiagen kit
• Protein Expression Est13
• Protein Expression FsC
• Western blot

Comamonas testosteroni

• Reviving a freez dried cell culture

In vitro

Nucleic acids

• Agarose gel electrophoresis
• PCR techniques
• SOE-PCR

DNA

• Agarose gel electrophoresis
• Ammonium acetate precipitation
• Cloning
• Dephosphorylation
• DNA Ligation
• DNA Ligation à la group transport
• DNA Quantification / NanoDrop
• Miniprep
• Mutagenic PCR
• PCR
• Purification of DNA
• Restriction digest
• Salting Out
• Sequencing DNA

Protein

• Activity assay
• Affinity chromatography
• Immobilized metal ion affinity chromatography (IMAC)
• pNP-Assay Bacteria
• pNP-Assay Enzyme
• Proteinpurification
• Purification of Periplasmatic Proteins
• SDS-Page