MOB-31812 Toolbox Molecular Biology

Course

Credits 12.00

Teaching methodContact hours
Thesis
Course coordinator(s)dr. HGJM Franssen
Lecturer(s)prof. dr. T Bisseling
dr. HGJM Franssen
dr. ir. R Geurts
dr. O Kulikova
dr. ir. EHM Limpens
JGJ Hontelez
Examiner(s)prof. dr. T Bisseling

Language of instruction:

Dutch and/or English

Assumed knowledge on:

Mob-20306 Gene Technology and MOB-30306 Control of Cellular Processes and Cell Differentiation or MOB-30806 Regulation of Plant Development or MOB-31303 Molecular Development

Contents:

Applications of DNA recombinant technology to biology have brought about a revolution in our understanding of living organisms. There is no field of fundamental and applied biological research that is untouched by the power we now have to isolate, analyze and manipulate genes. Whether taxonomy or gene therapy is in question - to mention the extremes - all research fields benefit from the potential of identifying, isolating and modifying genes. Because recombinant DNA techniques are instrumental to such a wide variety of research fields, it is of major importance for students of various disciplines to master various techniques and to acquire the molecular biological line of thought.
The primary goal of the MolBi-ToolBox is to provide students a broad, experimental overview of the diverse techniques for isolating, analyzing and manipulating genetic material. In concert, bioinformatics is applied either to extract data from databases or to do in silico experiments to analyse gene expression and gene function.
It is aimed to deal with a selection (based on the subject that will be worked on) of the following topics:
1. Isolation and physical mapping of genes
(i) Gene cloning / library screening
- genomic libraries;
- cDNA libraries;
- BAC/YAC libraries;
- PCR techniques.
(ii) Physical mapping
- Southern analysis/hybridization;
- Construction physical maps genomic and cDNA clones;
- Long-range mapping by PFGE.
2. Cyto-physical mapping of genomic and cDNA clones
Fluorescence in situ hybridization (FISH) on pachytene chromosomes;
Fibre-FISH on extended DNA fibres.
3. DNA sequencing and computer-directed genomic database screening Automated DNA sequencing;
Genomic DNA database handling.
4. Gene expression
- Differential RNA display;
- RT-PCR;
- Whole mount in situ hybridization;
- Expression vectors;
- Transient expression assays using reporter genes.
5. Gene manipulation and transfection
- Site-specific mutagenesis;
- GFP-fusions;
- Transfection of protoplast;
- Immunostaining.
6. Analysis transgenic cells
- Plasmid rescue sequences flanking T-DNA inserts (Arabidopsis);
- Cytogenetic mapping of transgenes (T-DNA inserts) by FISH.
7. In silico technologies
- database searches.
Due to a limited number of places available in the laboratory, students have to register at the secretariat of Molecular Biology and are encouraged to register in time. Students for which this course is compulsory or restricted optional take precedence.

Learning outcomes:

At the end of the course, the student is expected:
- to have a broad, experimental overview of the diverse techniques for isolating, analyzing and manipulating genetic material;
- to be able to collect background information relevant to the research topic;
- to be able to be carry out independently several experiments simultaneously;
- to be able to collaborate with companion students both in experiments as well as in documentation of the experiments;
- to be able to interpret the results of these experiments, and to incorporate the results in an oral presentation and a written report.

Activities:

- to do experiments according to the English written manuals and design experiments;
- to moderate with companion students;
- learn how to perform literature and data base searches;
- to work as a team, both in experiments as in writing a report;
- oral presentation.

Examination:

The final mark will be based on theoretical insight in the project and laboratory skills shown during the work, and on the quality of the oral presentation and written report about the results of the work.

Literature:

Literature is not available.

ProgrammePhaseSpecializationPeriod
Restricted Optional for: MBTBiotechnologyMScC: Medical Biotechnology1WD, 2WD, 6WD
MMLMolecular Life SciencesMSc1WD, 2WD, 6WD