Download e-book for kindle: Bioengineering for Pollution Prevention by John G. Taylor

By John G. Taylor

ISBN-10: 1606929003

ISBN-13: 9781606929001

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In this approach, the gene of interest has been identified and cloned, and PCR primers are designed for it. During amplification of the gene, however, a number of changes from normal PCR are employed: primer annealing temperatures are lowered to diminish fidelity; nucleotide ratios may be lowered and/or altered from the normal 1:1:1:1 equivalence; a nonproofreading DNA polymerase is used; high levels of the magnesium (II) ion (Mg2+) and often the manganese (II) ion (Mn2+) are used to further diminish replication fidelity; and up to 80 cycles may be conducted [21–23].

Because genes are expressed through the action of numerous enzymes that are conserved to some extent among organisms, it is often possible to express a cloned gene from one organism in another; moreover, it is also often possible to control the extent of expression very closely, as described below. 2 Genomics The field of genomics–the study of the sequence, structure, and function of an organism’s complete set of genetic information (its genome) [2]–is expanding rapidly due to the invention of numerous high-efficiency and high-throughput technologies capable of handling large amounts of DNA as well as sequence data.

Chemical and physical mutagenesis methods are relatively straightforward and inexpensive; however, additional methods have been sought because these methods typically affect only a few nucleotides at a time and therefore result in limited improvements [20]. 2. Error-prone PCR. Error-prone PCR is widely used to generate random mutants. In this approach, the gene of interest has been identified and cloned, and PCR primers are designed for it. During amplification of the gene, however, a number of changes from normal PCR are employed: primer annealing temperatures are lowered to diminish fidelity; nucleotide ratios may be lowered and/or altered from the normal 1:1:1:1 equivalence; a nonproofreading DNA polymerase is used; high levels of the magnesium (II) ion (Mg2+) and often the manganese (II) ion (Mn2+) are used to further diminish replication fidelity; and up to 80 cycles may be conducted [21–23].

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Bioengineering for Pollution Prevention by John G. Taylor


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