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Laboratory Help Upper-Year Courses Topic started by: gkor29 on Mar 17, 2015



Title: Design an experiment outlining your techniques that will analyze the temporal expression pattern of
Post by: gkor29 on Mar 17, 2015
Design an experiment outlining your techniques that will analyze the temporal expression pattern of a gene (gene X - mRNA expression) and its protein localization during gastrulation in three different organisms.
Please state the organisms you are choosing, the time in which you will perform the experiment.
Please also state the molecular techniques you will use to follow gene expression (mRNA and protein)
Marks will be based on the accuracy and feasibility of your design, and the important scientific details of the experiment.

I'm a little unsure about how to format this experiment and whether or not the there organisms used should correlate to each other. From what i understand a friend of mine took a different approach and i think it confused me more than helped me

Friends example:
Experiment 1: (Determining Head Development in Drosophilia melanogaster)In this experiment I will be using a reporter gene and attach it to the bicoid gene of a fruit fly via a plasmid. Since I am testing for the temporal expression of this gene during different stages of development, this seems like a reasonable technique being how it won't affect the expression of the gene,. It will also serve as a marker for the successful uptake of the gene. In order to accurately observe the expression of this gene I will be tagging the gene with a fluorescent protein in order to visualize it under UV-light. By testing the concentration gradients of this gene during the different stages of gastrulation, I am able to see how it's expression get's regulated as the organism develops.
Experiment 2: (Testing how temperature affects color in Siamese cats)The Himalayan gene is temperature sensitive in Siamese cats which will determine what colors will be expressed in specific areas on Siamese ! cats. A technique to track this genes throughout the develop, northern blot testing can be used by exposing a cell with a protease. From here I can separate the mRNA by gel electrophoresis and transfer it through a filter by blotting. After this step, I would incubate the gene with a probe and test the bands that appear to tell how much of this gene is being expressed.
Experiment 3: (Finding facial malformations in human)By using microarray technology, I was able to identify genes that allow for the specification of facial features in humans. By taking samples from an embryo through different stages of development I was able to amplify the genes to test for different levels of expression of facial features. Through the induction of mercury into the embryo, I was able to compared the genes in both healthy embryos and toxic laden embryos, and then finally compare how they manifest as it develops.


Thank you ahead of time