Cloning strategies

Target Group

Technicians and researchers with previous knowledge in molecular biology who want to get a comprehensive overview of cloning strategies and want to optimize their cloning protocols

Content/Learning Objective

Within this one-and-a-half-day theory course, basic methods and strategies for cloning of DNA fragments are presented. Topics covered include:

• cloning strategies:
  vectors: possibilities and choice (which vectors are available for which applications: standard cloning vectors, eukaryotic expression vectors, inducible systems etc.)
  enzymes: isochizomeres, compatible ends, suitable standard enzymes, fill-in and blunt end, dephosphorylation etc. Tricks and help for tricky cloning
• DNA quality and purity requirements: gel purification and isolation
• ligation: stoichiometrie and discussion of different methods
• transformation: competent bacteria, generation and efficiency control
• selection: resistance to antibiotics, blue-white-selection using ß-galactosidase genes
• analysis and control of results
• trouble shooting

Main Topics

Strategies for successful and efficient cloning as well as multiplication of DNA fragments.