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Bouffées de chaleur avec rougeur circonscritedes joues, sensation de brûlures intense des paumes des mains, des plantes de pieds et des oreilles
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- 10.23 Sara is an undergraduate student who is doing an internship in the research laboratory described in Questions 10.14 and 10.22. Just before Sara started working in the lab, the restriction map in Figure 10.D was made of the 47-kb NotI restriction fragment containing the prion protein gene (distances between restriction sites are in kb). Since smaller DNA fragments cloned into plasmids are more easily analyzed than large DNA fragments cloned Table hereinto BACs, Sara has been asked to "subclone" the 6.1-, 10.5-, 4.1- and 8.2-kb BamHI DNA fragments containing the prion-protein gene into the pBluescript II plasmid vector (see Figure 8.4, p. 174, for a description of pBluescript II). Her mentor gives her some intactpBluescript II plasmid DNA, some of the purified 47-kb NotI fragment, and shows her where the stocks of DNA ligase, BamHI, and reagents for PCR are stored inthe lab.a. Describe the steps Sara should take to complete her task if she has no information about the sequence of the 47-kb NotI fragment. In your answer, address how she will identify plasmids that contain genomic DNA inserts, and how she will verify that she has identified clones containing each of the desired genomic BamHIfragments.b. Describe an alternative approach that Sara could taketo complete her task if she first performs a bioinformatic analysis utilizing DNA sequence information available from the mouse genome project, and identifies the sequence of the 47-kb NotI fragment