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建立人际资源圈Genetics_Lab
2013-11-13 来源: 类别: 更多范文
Genetics Lab
Genetics Lab
Introduction:
The objective of this lab is to prepare a genomic library that will be later screened to obtain a clone that contains (LUX) operon of the studied bacterium vibrio fischeri. This requires several steps including DNA isolation, restriction digestion, electrophoresis and then ligation. The results from the experiments above can be used in many interesting ways, not only to clone known genes, but also to disclose the limitless amount of undiscovered genes that have potential in medicine and gene therapy.
Also, it presents a system that allows performing the isolation of the genomic library of bacterium vibrio fischeri clone and preparation and study of the plasmid subclone in a short period of time.
Isolation of plasmid DNA involves: growth of bacteria and amplification of plasmid, harvesting and lysis of bacteria, and purification of plasmid DNA.
DNA isolation from a particular organism requires celll lysis. Lysozyme enzyme is used to allow access to peptidoglycan and degrade it so that isolation of DNA can be facilitated. The most important reagents used in DNA isolation are: phenol and chloroform that dissociate protein from nucleic acid. Ethanol is very important to precipitate DNA provided and ionic strength of the solution is high and DNases are
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