Practical Course on Rapid Cloning, Protein Expression, and Purification CPEP 18 - 3 ECTS

This course is organised in the collaboration with the Protein Expertise Platform (PEP). It is based on the EMBO PEPC5 course.

Date: Wednesday 28th February to Friday 9th March 2018
Start: Wednesday, 28th February 2018, 12:30
END: Friday, 9 March 2018, 12:00

Place: Biochemistry teaching lab, KB.F4.04, KBC-building 4th floor (2 stairs up), Linnaeus väg 6, Umeå University

Time: About 6-8 h per day, mainly laboratory work but also lectures, discussions & seminars
Course leader: Uwe Sauer, KBC, 786-5930, E-mail: uwe.sauer@umu.se    Instructor: Gunter Stier, Heidelberg Univ., Germany, E-mail: gunter.stier@bzh.uni-heidelberg.de  

EXTENDED Registration deadline: Friday, 16th of February, 2018
(Admission on a first-come first-served basis)

The registration is closed now!

Participants: max. 12 (PhD students, post-docs or technical staff)

NO Participation fee!

Credits: 3 ECTS on a pass – no pass basis. Full time participation is required to obtain a pass certificate.

More information about the course:http://babel.ucmp.umu.se/cpep
Aims of the practical CPEP course:
After the course you should know how to:

  • Plan and successfully execute your project. Bring your own project(s)!(*)
  • Make efficient use of bioinformatics tools for planning and analysis of protein expression projects
  • Design PCR primers and codon optimize genes
  • Apply various cloning and mutagenesis techniques
  • Use medium throughput approaches for cloning and protein expression in E. coli
  • Carry out fast parallel cloning of your gene(s) into vectors containing different tags and fusion proteins
  • Use of fusion proteins and associated purification strategies
  • Test for optimal protein production: different strains, growth conditions and fusion partners
  • Co-express components of protein complexes and their purification strategies
  • Prepare and use of auto-inducing medium
  • Do protein expression mini-screens
  • Scale-up positive screening candidates to a protein midi-preparation using purification machines
  • Fc-fusion protein expression in mammalian cells
  • Trouble shoot problems and apply strategies to solve them
  • Apply special protein labeling techniques, such as SeMet, 15N-labeling for quick NMR analysis

Additional demonstrations will include the use of:

  • Protein purification with “Äkta” machines
  • The Mosquito crystallization robot, crystal imaging facility
  • Crystallization and X-ray diffraction of crystals and analysis of diffraction data
  • NMR characterization of 15N labeled proteins and interpretation of simple NMR spectra
  • EM facility – first approach to sample preparation (negative staining) and electron microscopy measurements

Practical sessions are complemented with lectures, round-table discussions about techniques, problems and their solutions. Participants will present short seminar talks of selected topics.

(*) If you bring your own project(s), you should contact the instructors at least 2 weeks before the start of the course (preferably no later than February 16, 2018).

The course leaders will provide you with a project if you do not bring one.


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