Toolbox Molecular Biology

MolBi-Toolbox

Registration

Three times a year you can participate in the ToolBox, in the 1st, 2nd, and 6th period respectively. Due to the intensive supervision of the ToolBox the number of participants is restricted to 6 maximum.

Therefore early birds sign in at the secretariat of the Laboratory of Molecular Biology. Please, do not register for ToolBox using EDUweb but contact our secretariat.

Aims

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. Furthermore, the students are expected to collect background information themselves relevant to the research they are involved in. The student will learn to be involved in several experiments simultaneously. Collaboration with companion students both in experiments as well as in documentation of the experiments is a prerequisite for a successful Toolbox.

Mandatory knowledge

MOB-20306 Gene Technology, and one of the following courses: MOB-30306 Control of Cellular Processes and Cell Differentiation, MOB-30806 Regulation of Plant Development or MOB-31303 Molecular Development, or equivalent.

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.
As stated, 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.

Based on the subject that will be worked on, it is aimed to deal with a selection of the following topics:

  1. Isolation and physical mapping of genes
    • Gene clining/ library screening
      • genomic libraries
      • cDNA libraries
      • BAC/YAC libraries
      • PCR techniques
    • Physical mapping
      • Southern analysis/hybridization
      • Construction physcal maps genomic and cDNA clones
      • long-rane 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