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Who made the first successful airplane flight?

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*10.28 Katrina purified a clone from a plasmid library made using genomic DNA and sequenced a 500-bp long segment using the dideoxy sequencing method. Hertwin-sister Marina used PCR with Taq DNA polymerase to amplify the same 500-bp fragment from genomic DNA. Marina sequenced the fragment using the dideoxysequencing method, and obtained the same sequence as Katrina did. She then cloned the fragment into a plasmid vector and, following ligation and transformation into E.coli, sequenced several, independently isolated plasmidsto verify that she had cloned the correct sequence. Most of them have the same sequence as Katrina's clone, but Marina finds that about 1/3 of them have a sequence that differs in one or two base-pairs. None of the clones that differ from Katrina's clone are identical. Fearing she has done something wrong, Marina repeats her work, only to obtain the same results: about 1/3 of the fragments cloned from the PCR product have single base-pair differences. Explain this discrepancy.

*10.30 Chapter 9 presented a description of how DNA microarray analysis was used to characterize changes to the transcriptome during yeast sporulation. That analysis found that more than 1,000 yeast genes showed significant changes in mRNA levels during sporulation, identifiedat least seven distinct temporal patterns of gene induction, and provided insights into the functions of many orphan genes. It is important to confirm findingsfrom microarray analyses using independent methods. How would you confirm independently that three orphan genes display altered expression during yeast sporulation?

*10.24 Imagine that you have cloned the structural gene for an enzyme that functions in the biosynthesis of catecholaminesin the adrenal gland of rats. How could you use this cloned DNA as a probe to determine whether this same gene functions in the rat brain?

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