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Unc-119 Gene Lab Report

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A forward genetics screen will be performed for unc-119 C. elegans. The experiment will be conducted based on visible or observable phenotypes using a dissecting microscope. F2 population will be observed for abnormal movement phenotype on a solid medium. It is known that with uncoordinated mutations the organism has no sense of direction, the organism tend to move back and forth only in comparison to a control (normal) organism. Different trial plates will be performed for better results. In this part of the experiment the F2 generation will be studied to identify the defects of unc-119 gene, and how it affects the living cycle of the organism. The organisms that show mutation will be isolated and visually screened. F3 generation will be analyzed to determine if they show the same phenotype or what type of mutations.
Reverse Genetic Screen: …show more content…
A plasmid will be created with the transcriptional control for the gene of interest connected to a reporter gene. Then, the plasmid created would be injected into the organism and will allow the reported gene to be expressed. A reported gene used will be ß-galactosidase because this enzyme will show a blue stain when mutated gene is being expressed. One worm will be strain with the plasmid that contains the promoter from unc-119 gene fused to lacZ, and another worm (control) will be strain with a plasmid that contains a promoter from another mutated gene. This will help determine the expression of the gene of interest by staining it. The worms will be micro centrifuged, and then stained with the blue dye. Then will be allowed to dry until the blue stain shows. The slides with the C. elegans will be analyzed under a dissecting microscope to determine the expression of the gene. If it fluoresces, then it means the gene its being expressed and the phenotype will be observed (Jones). Protocol by Jones, D., et

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