Methods in Molecular Biology – Theory and Practice
Background
Atherosclerosis is a progressive inflammatory disease. Macrophages play a dominating role in
progression of atherosclerosis, as they remove, internalize and metabolize retained
lipoproteins, giving rise to cells known as lipid-laden "foam cells". These foam cells promote
atherogenesis by a number of mechanisms, including secretion of inflammatory cytokines,
and although they have a long half-life, lesional macrophages undergo a steady rate of
apoptosis. These foam cells and other dying inflammatory cells are cleared by neighboring
macrophages. Therefore, the ability to maintain effective clearance of foam cells is necessary
to limit atherosclerosis progression. One novel molecule that may influence macrophage
function is CD93. This molecule is expressed on macrophages and upon stimulation is shed
from the surface. Interestingly there are genetic variants of this molecule, which may play a
role in atherosclerosis development. Some of the basic methods to study candidate genes that
contribute to atherosclerosis include genotyping, real time RT-PCR, flow cytometry and
ELISA to detect SNP, RNA and protein expression respectively.
Contents of the lab
The themes of the lab are:
- Practical and theoretical review of basic methods in molecular biology such as:
leukocyte isolation, RNA and DNA extraction, gene expression and genotype
determination, flow cytometry and ELISA
- Introduction to basic statistical methods for hypothesis testing
- Collection, presentation and discussion of scientific results
Learning outcomes
By the end of the course the students are expected to have:
- Increased practical and theoretical knowledge of basic methods in molecular biology
- Increased understanding of how to collect and present scientific results
- Increased understanding of why statistics are needed in science and which basic
hypothesis testing methods are commonly used.
- Gained experience in writing a scientific report
Examination
Examination of the learning outcomes is achieved by:
1) Writing of a short individual report of the laboratory experiment (max 1 A4 page, 12
point, Times New Roman). The report should be written in abstract form with the
headings: Title, Background, Aims, Methods, Results, Conclusions.
2) Peer review and discussion of other student’s laboratory reports.
Compulsory parts
100% attendance of the laboratory parts and the group discussion.
Possibilities to compensate for absence can be discussed with the lab organisers.
Grading
Pass / Fail.
To pass the lab, 100% attendance of the compulsory parts and fulfilment of the examination
assignments are required (see above).
Literature
Compendium of molecular biology methods (will be handed out at the beginning of the
course) and recommended articles.
Lab organisers
Josefin Skogsberg josefin.skogsberg@ki.se
Dick Wågsäter
dick.wagsater@ki.se
070-714 23 01
076-839 73 99