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From 2013.igem.org

Labbook |	Protocols |	Materials |

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

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

In vivo

Escherichia coli

• Bacterial cell culture
• Bacterial transformation
• Cell counting/plating
• Chemically competent cells
• Colony PCR
• Glycerine Stocks
• Heat Shock Transformation
• PureYield Plasmid Miniprep System
• Expression of LssmOrange and mKate

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