les1 - INTRODUCTION Case 1: Your supervisor has heard at a...

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1 INTRODUCTION Case 1 : Your supervisor has heard at a congress about this very interesting promising new gene of which only the name is known. He wants you to write a project on the protein and start to perform experiments. After a literature search you only know that the encoded protein upon overexpression is involved in the activation of a certain transcription factor. Case 2 : Suppose a biotech company recruits you. Your job is to develop and optimize high through put cell-based assays to screen drug databases for drugs that inhibit inflammation. How will you tackle these problems? What are the logical steps to follow? Which techniques will you use? How will you design your experiment? In order to find a good strategy to be able to resolve problems like these, a profound knowledge of molecular biomedical techniques is indispensable. With the course of Experimental Molecular Biology, we want to -Provide some advanced biochemical, molecular and cell biological techniques that are commonly used in modern biomedical research. -Provide those who anticipate careers involving biological or medical research with sufficient background information so that they are able to design experiments and know which methods are best suited for a particular research question or problem in the given research environment. -Provide students a better access to the scientific literature in that understanding the methods will allow students to critically evaluate the results and conclusions of scientific papers.
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2 Lectures will describe the theories and principles behind each of the methods in addition to discussing the practical aspects and limitations in executing the various procedures. Coming back to the two cases, how to start with your research? 1. Scientific literature search : Before starting your research, you have to get to know the published data, which you may find at the following URL: http://www.ncbi.nlm.nih.gov/sites/entrez 2. Bio-informatic tools: some useful URL’s : http://www.ncbi.nlm.nih.gov/sites/entrez ExPASy Molecular Biology server: http://www.expasy.ch For more information concerning bio-informatics, we refer to the course “Introduction in bio-informatics” (3 th Bachelor) which deals with these aspects 3. Setup of your experiments : Some general guidelines: - Choose your model organism, type of tissue, cells: tissue culture (Chapter I) - Choose the right conditions (e.g. stimulus, time kinetics, transfection methods…) and the right biochemical, molecular and cell biological technique(s) that will potentially give you the best answer to your question. Keep in mind the limitations of the techniques, alternatives, … - Choose the right positive and negative controls - Perform your experiment in the tissue culture accurately conform GLP - Once your experiment is finished in the tissue culture, you can proceed in different ways dependent on the goal of your experiment: you can leave your cells intact (e.g. for ex vivo studies e.g. sorting of cells), fix them (e.g. on cover
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les1 - INTRODUCTION Case 1: Your supervisor has heard at a...

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