Methods

Here are links to some of the methods used and resources avaliable in our group.

Feel free to use. If useful, thanks are always gratefully accepted!

• Cloning an expression construct (aka “The vintage manual of cloning”)
• A more modern version for SLIC cloning
• Testing for expression and solubility
• Large scale expression with pBAT vectors
• Protein quantification with UV absorbance (updated 2017)
• SDS-PAGE
• Zinc-Imidazole staining for SDS-PAGE gels
• Inclusion body preparation
• Western blotting

You might also find useful the following cloning, expression etc. related materials:

• E. coli expression vectors (all available now from Addgene!)
  • pBAT series of T7 expression vectors we have used for years
  • The improved version of these vectors, our new pOP series of plasmids
  • Vectors with a wider range of fusions and for SLIC: the pExp series of vectors

Some slides from methods talks

• Slides from 2022 postgraduate course on protein production at University of São Paulo