Integrated practicum: molecular biology and genetics

Course Code :1062FBDBMW
Study domain:Biomedical Sciences
Academic year:2017-2018
Semester:1st semester
Contact hours:60
Study load (hours):84
Contract restrictions: Exam contract not possible
Language of instruction:Dutch
Exam period:exam in the 1st semester
Lecturer(s)Eva Geuens
Guy Van Camp

3. Course contents *

The practicum is divided into five major experiments:

  • Experiment I: Sequencing of an unknown gene. In this experiment, sequencing of an unknown gene is performed on the students own DNA extracted from saliva. During this the following techniques are used: DNA isolation, agarose gel electrophoresis, DNA amplification using polymerase chain reaction (PCR), ligation of a DNA fragment in a plasmid vector, heat shock transformation, Lac-Z screening, plasmid DNA extraction and DNA sequencing.
  • Experiment II: Genotyping of polymorphic DNA markers - segregation analysis. In the second experiment, the students’ DNA extracted from saliva (Experiment 1), together with DNA of a family will to be analysed using polymorphic genetic markers followed by segregation analysis in order to construct the pedigree of the family.
  • Experiment III: SNP detection using High Resolution Melting analysis. During this experiment, the zygosity of the SNP rs3747174 in the CBY gene will be evaluated using HRM-analysis on students’ DNA extracted from saliva and DNA of different controls persons.
  • Experiment IV: Q-PCR using SYBRGreen. During this experiment, the difference in expression of the APP-23 gene in the brain tissue is quantitatively evaluated in a control and a APP overexpression mouse.
  • Experiment V: Mutation detection by RFPL - PCR. In this experiment, the DNA of a number of controls and individuals with a given condition is analyzed by RFLP-PCR to determine which persons are carrying the mutation and which are not. Specifically, a mutation resulting in the formation of the restriction site for the restriction enzyme HindIII is evaluated.