By Dr Desmond S. T. Nicholl

ISBN-10: 0521004713

ISBN-13: 9780521004718

Des Nicholl offers a brand new, absolutely revised, and improved version of his well known undergraduate-level textbook. The e-book keeps a number of the positive factors of the unique variation and nonetheless deals a concise technical advent to the topic of genetic engineering. it's divided into 3 major sections: uncomplicated molecular biology, equipment of gene manipulation, and sleek functions of genetic engineering. functions lined within the booklet contain genomics, protein engineering, gene treatment, cloning, transgenic animals and vegetation, and bioethics. An advent to Genetic Engineering is vital analyzing for undergraduate scholars of biotechnology, genetics, molecular biology, and biochemistry.

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The nucleic acids in the gel are usually visualised by staining with the intercalating dye ethidium bromide and examining under UV light. Nucleic acids show up as orange bands, which can be photographed to provide a record (Fig. 6). The data can be used to estimate the sizes of unknown fragments by construction of a calibration curve using standards of known size, as migration is inversely proportional to the log10 of the number of base-pairs. 3). In addition to its use in the analysis of nucleic acids, polyacrylamide gel electrophoresis (PAGE) is used extensively for the analysis of proteins.

These dNTPs lack a hydroxyl group at the 3Ј position of deoxyribose, which is necessary for chain elongation to proceed. Such modified dNTPs are known as dideoxynucleoside triphosphates (ddNTPs). The four ddNTPs (A,G,T and C forms) are included in a series of four reactions, each of which contains the four normal dNTPs. The concentration of the dideoxy form is such that it will be incorporated into the growing DNA chain infrequently. Each reaction therefore produces a series of fragments terminating at a specific nucleotide, and the four reactions together provide a set of nested fragments.

The DNA to be probed is also denatured, and is usually fixed to a supporting membrane made from nitrocellulose or nylon. Hybridisation is carried out in a sealed plastic bag or tube at 65–68 °C for several hours to allow the duplexes to form. The excess probe is then washed off and the degree of hybridisation can be monitored by counting the sample in a scintillation spectrometer or preparing an autoradiogram, where the sample is exposed to X-ray film. Some of the applications of nucleic acid hybridisation as a method for identifying cloned DNA fragments will be discussed in Chapter 8.

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