werkcollege_090310-final

werkcollege_090310-final - Work college I - 090310 1...

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Unformatted text preview: Work college I - 090310 1 EXPERIMENTAL MOLECULAR CELL BIOLOGY WORK COLLEGE I 10/03/2009 Work college I - 090310 2 1. Introduction 1.1. Part 1 The first intention of this work college is to implement the molecular cell biology techniques that have been extensively discussed in the lectures. For this purpose, we have drafted questions build around a certain signalling pathway. In these questions, we ask you to think thoroughly over a strategy using one or more of the techniques from the course that you would use to resolve the problem. These type of exercises will help you in the future to think out experiments of your own e.g. during your master thesis. It might also elucidate some of the techniques that have been processed in the lectures. How? Write out the experiment you propose step by step, how you would perform the experiment in the lab, meaning e.g.: 1. Which construct will you make, do you need to transfect cells, … 2. Which are the -/+ controls that you can use (if possible),… 3. Are you going to stimulate your cells, perform a time kinetic experiment, … 4. Which technique would you use? There may be several possibilities, but try to argument your choice! 5. Are you going to lyse your cells, is it an in vivo or an in vitro exp,… 6. What is the read-out of your experiment, what is the detection method,… This type of questions can be found back in part 1, question 1 till 10 . 1.2. Part 2 The second goal of this work college is to learn to interpret data, figures or tables originating from scientific articles or life science company websites. We ask you to think about the following questions: 1. Which technique has been used in the experiment? Interpret the data. 2. Propose an alternative strategy to evaluate the same question. These questions deal with part 2, question 1 till 9 . These types of questions are comparable to the exam questions! Work college I - 090310 3 1.3. Organization • 19 groups of 2 students • Each group resolves 3 questions (2 questions on topic 1; 1 questions on topic 2) (see table) • preparation time: 14h - 15h • Spare time: start with the other questions = beneficial for you! • Each group reports orally on 1 of the prepared questions: – for part 1: • student A reports on sub-questions 1 till 3 • student B reports on sub-questions 4 till 6 – for part 2: • student A reports on sub-question 1 • student B reports on sub-question 2 Student Group I II Evaluation A B Q Q Q A B 1 1 2 1 2 3 4 2 3 5 6 3 4 7 8 4 5 9 10 5 6 1 2 6 7 3 4 7 8 5 6 8 9 7 8 9 10 9 10 1 11 1 2 1 12 3 4 2 13 5 6 3 14 7 8 4 15 9 10 5 16 1 2 6 17 3 4 7 18 5 6 8 19 7 8 9 Work college I - 090310 4 1.4. Part 1: Thinking out experiments Figure 1. TLR4 signalling pathway . Following stimulation with LPS (lipopolysaccharide) and dimerisation, the TLR4 signalling pathway recruits the adaptor molecule MyD88 and induces nuclear factor (NF)- κ B and mitogen-activated protein kinases (MAPKs) through TLR4 associated kinase (IRAK)-4, IRAK-1 and TNF receptor associated factor (TRAF)-6. In addition, a MyD88-kinase (IRAK)-4, IRAK-1 and TNF receptor associated factor (TRAF)-6....
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werkcollege_090310-final - Work college I - 090310 1...

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