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Preparation of oligo solutions

PCR experiments

Digest of insert DNA

Digest and dephosphorylation of vector DNA

Ligation of DNA fragments

Preparation of chemically competent cells

Transformation of plasmid DNA into chemically competent cells

Ligase independent cloning (LIC)

Ligase independent cloning (LIC) using pETM-13/LIC

• Creating a construct with a C-terminal His6-tag

Ligase independent cloning (LIC) protocols

• Ligase independent cloning (LIC)
• Ligase independent cloning (LIC) using pETM-13/LIC
  Creating a construct with a C-terminal His6-tag

Culture media

Auto-induction media

• Studier, F.W. (2005) Protein Expression and Purification 41, 207-234

M9 Minimal medium

Lysogeny broth (LB)

Terrific Broth (TB)

Other bacterial media

Fermentation protocols

High cell-density fermentation in E. coli

Protein production protocols

Production of His-tagged TEV protease

Production of His-tagged 3C protease 

Production of GST-tagged 3C protease

Biophysical characterization of proteins

Static light scattering

• Nettleship, J.E., Brown J., Groves M.R., Geerlof A. (2008) Methods Mol. Biol. 426, 299-318.

Isotope labelling of proteins

• ¹⁵N-labelling of proteins in E. coli
• ¹³C-labelling of proteins in E. coli
• ¹⁵N and ¹³C-labelling of proteins in E. coli