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James Joyce used this literary technique to present characters' thoughts and feelings?

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*10.20 During Southern blot analysis, DNA is separated by size using gel electrophoresis, and then transferred to a membrane filter. Before it is transferred, the gel issoaked in an alkaline solution to denature the doublestrandedDNA, and then neutralized. Why is it important to denature the double-stranded DNA? (Hint: Consider how the membrane will be probed.)

10.31 Metalloproteases are enzymes that require a metalion as a cofactor when they cleave peptide bonds. Members of one family of metalloproteases share the following consensus amino acid sequence in their catalytic site:His-Glu-X-Gly-His-Asp-X-Gly-X-X-His-Asp (X is any amino acid). Structural models of the catalytic site developed from X-ray crystallographic data suggest that the second amino acid, glutamate, is essential for proteolytic activity.Outline the experimental steps you would take to test this hypothesis. Assume you possess a cDNA encoding a metalloprotease having the consensus sequence, and canmeasure metalloprotease activity in a biochemical assay.

*10.21 A researcher digests genomic DNA with the restrictionenzyme EcoRI, separates it by size on an agarose gel, and transfers the DNA fragments in the gel to a membrane filter using the Southern blot procedure. What result would she expect to see if the source of the DNA and the probe for the blot is described as follows?a. The genomic DNA is from a normal human. The probe is a 2.0-kb DNA fragment excised by the enzyme EcoRI from a plasmid containing single-copy genomic DNA.b. The genomic DNA is from a normal human. The probe is a 5.0-kb DNA fragment that is a copy of a LINE ("long interspersed element", a type of repetitive sequence; see Chapter 2, p. 29 and Chapter 7, pp. 160-161) that has an internal EcoRI site.c. The genomic DNA is from a normal human. The probe is a 5.0-kb DNA fragment that is a copy of a LINE that lacks an internal EcoRI site.d. The genomic DNA is from a human heterozygous for a translocation (exchange of chromosome parts) betweenchromosomes 14 and 21. The probe is a 3.0-kb DNA fragment that is obtained by excision with the enzyme EcoRI from a plasmid containing single-copy genomic DNA from a normal chromosome 14. The translocation breakpoint on chromosome 14 lies within the 3.0-kb genomic DNA fragment.e. The genomic DNA is from a normal female. The probe is a 5.0-kb DNA fragment containing part of the testis determining factor TDF gene, a gene located on the Y chromosome.

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