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By Dr Desmond S. T. Nicholl

Des Nicholl offers a brand new, totally revised, and elevated version of his well known undergraduate-level textbook. The publication keeps the various positive aspects of the unique version and nonetheless deals a concise technical advent to the topic of genetic engineering. it really is divided into 3 major sections: simple molecular biology, equipment of gene manipulation, and sleek purposes of genetic engineering. purposes coated within the e-book contain genomics, protein engineering, gene treatment, cloning, transgenic animals and vegetation and bioethics. An advent to Genetic Engineering is key studying for undergraduate scholars of biotechnology, genetics, molecular biology, and biochemistry.

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A) Nuclease Bal 31 is a complex enzyme. Its primary activity is a fast-acting 3Ј exonuclease, which is coupled with a slow-acting endonuclease. When Bal 31 is present at a high concentration these activities effectively shorten DNA molecules from both termini. (b) Exonuclease III is a 3Ј exonuclease that generates molecules with protruding 5Ј termini. (c) DNase I cuts either single-stranded or double-stranded DNA at essentially random sites. (d) Nuclease S1 is specific for single-stranded RNA or DNA.

2. This illustrates the tremendous range of sizes for human genes, the smallest of which may be only a few hundred base-pairs in length. 2. 6 Note: Gene sizes are given in kilobase-pairs (kbp). The number of exons is shown, and the percentage of the gene that is represented by these exons is given in the final column. Fig. 7. Structure and expression of the mammalian ␤-globin gene. The gene contains two intervening sequences or introns. The expressed sequences (exons) are shaded and numbered. The primary transcript is processed by capping, polyadenylation and splicing to yield the fully functional mRNA.

The technique of gradient centrifugation is often used to prepare DNA, particularly plasmid DNA (pDNA). In this technique a caesium chloride solution containing the DNA preparation is spun at high speed in an ultracentrifuge. Over a long period (up to 48 h in some cases) a density gradient is formed and the pDNA forms a band at one position in the centrifuge tube. The band may be taken off and the CsCl removed by dialysis to give a pure preparation of pDNA. As an alternative to gradient centrifugation, size exclusion chromatography (gel filtration) or similar techniques may be used.

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