Experimental biochemistry and biophysics
CH-319 / 4 credits
Teacher: Hovius Rudolf
Language: English
Withdrawal: It is not allowed to withdraw from this subject after the registration deadline.
Summary
A 7-week long (4+8 h) experiment where you plan and construct a fluorescent sensor protein starting from DNA bricks. The protein will be expressed in and purified from E.coli, characterized by biochemical and spectroscopic methods. Your report will be in the style of a scientific paper or poster
Content
i) Molecular biology: DNA fragment isolation by PCR, DNA restriction and ligation, plasmid purification and sequence analysis, agarose gel electrophoresis.
ii) Biochemistry: Bacterial protein expression, protein purification by affinity chromatography, analysis by SDS-PAGE and spectroscopy, fluorescent labelling
iii) Biophysics: Fluorescence spectroscopy, enzyme kinetics or molecular interactions.
Keywords
Molecular biology - cloning
Protein expression & purification
Bio-chemical and bio-physical chracterisation
Sensor proteins
Reporting
Learning Prerequisites
Required courses
Admission to the TP is conditional on the successful completion of 2 of the 3 courses: CH-210 Biochimie, CH-313 Chemical biology, CH-311 Macromolecular structure and interactions
Recommended courses
Molecular and cellular biophysics I (CH-311)
Chemical Biology (CH-313)
Important concepts to start the course
genetic engineering & DNA manipulation; protein synthesis; DNA & protein analysis; absorbance and fluorescence spectrometry; enzymology, molecular interactions
Learning Outcomes
- Design cloning strategy
- Produce a scientific report and high-quality lab journal
- Integrate Good laboratory behavior and wet lab practice
- Assess / Evaluate your data critically
- Produce a purified expressed protein
- Analyze proteins and DNA
- Characterize sensor function
- Use common sense and logical deduction
- Interpret protein structure data
- Design cloning strategy
- Produce a scientific report and high-quality lab journal
- Integrate Good laboratory behavior and wet lab practice
- Assess / Evaluate Evaluate your data critically
- Produce a purified expressed protein
- Analyze proteins and DNA
- Characterize sensor function
- Interpret protein structure data
Transversal skills
- Collect data.
- Continue to work through difficulties or initial failure to find optimal solutions.
- Write a scientific or technical report.
- Plan and carry out activities in a way which makes optimal use of available time and other resources.
Teaching methods
Students prepare and discuss experimental approach
Laboratory experimentation
Discussion of experimental progress and results
Reporting on scientific level
Topiocal presentations
Expected student activities
Good theoretical preparation & planning of lab work before doing experiments; skillful execution of experiments, being organized & keeping a lab book; thorough analysis of results, writting a scientific-grade report; respecting security rules and fellow students.
Assessment methods
valuation of preparation and planning
Evaluation of experimental execution, including good laboratory behaviour
Evaluation of comprehension through discussion & written questions
Evaluation of report: including structure, data treatment and presentation, critical attitude , comparison to scientific literature
Supervision
| Office hours | Yes |
| Assistants | Yes |
| Others |
Resources
Bibliography
Bibliography
TP manual
Biochemistry & Biophysics text books
ApE; a plasmid editor free software
Methods, Structures, and other useful info via Moodle
Notes/Handbook
Manual
Websites
Moodle Link
Prerequisite for
big plus for Msc courses and for semester or diploma projects in chemical biology or biophysics
In the programs
- Semester: Spring
- Exam form: During the semester (summer session)
- Subject examined: Experimental biochemistry and biophysics
- TP: 6 Hour(s) per week x 14 weeks
- Type: mandatory
- Semester: Spring
- Exam form: During the semester (summer session)
- Subject examined: Experimental biochemistry and biophysics
- TP: 6 Hour(s) per week x 14 weeks
- Type: optional