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what is the size of the ocean carbon store (Gt)?

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10.25 A cDNA library is made with mRNA isolated from liver tissue and the vector shown in Figure 10.4. When a cloned cDNA insert from that library is digested with the enzymes EcoRI (E), HindIII (H), and BamHI (B) (described in Table 8.1, p. 174), the restriction map shown in the following figure, part (a), is obtained. When this cDNA isused to screen a cDNA library made with mRNA frombrain tissue and the vector shown in Figure 10.4, three identical cDNAs with the restriction map shown in the following figure, part (b), are obtained. When a uniformlylabeled, 32P-labeled riboprobe made using T7 RNA polymeraseis prepared using either cDNA and the probe is allowed to hybridize to a Southern blot prepared fromgenomic DNA digested singly with the enzymes EcoRI, HindIII, and BamHI, an autoradiograph shows the pattern of bands in the following figure, part (c). When any of the -32P labeled riboprobes are used to probe a northern blotprepared with poly(A)+ mRNA isolated from liver and brain tissues, no signal is seen. However, when the samenorthern blot is probed with a uniformly labeled, -32P labeled probe is prepared using the random primer method (described in Box 10.1), the pattern of bands in part (d) of the figure is seen. Fully analyze these data and then answer the following questions:a. Do these cDNAs derive from the same gene?b. Why are different-sized bands seen on the northern blot?c. Why could hybridization signal be detected on the Southern blot but not on the northern blot when riboprobes were used? Why could hybridization signal be detected on the northern blot when a random-primed probe was used? (Hint: consider how these probes aremade and which nucleic acid strands become labeled.)d. Why do the cDNAs have different restriction maps?e. Why are some of the bands seen on the whole genome Southern blot different sizes than some of the restriction fragments in the cDNAs?

*10.34 The maps of the sites for restriction enzyme R in the wild type and the mutated cystic fibrosis genes are shown schematically in the following figure: Table hereSamples of DNA obtained from a fetus (F) and her parents (M and P) were analyzed by gel electrophoresis followed by the Southern blot technique and hybridization with the radioactively labeled probe designated "CF probe" in the previous figure. The autoradiographic results are shown in the following figure: Figure hereGiven that cystic fibrosis results from a recessive trait and affected individuals always have two mutant alleles, will the fetus be affected? Explain.

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

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