By Dr Desmond S. T. Nicholl
Des Nicholl offers a brand new, absolutely revised, and extended variation of his well known undergraduate-level textbook. The e-book keeps the various good points of the unique variation and nonetheless deals a concise technical advent to the topic of genetic engineering. it really is divided into 3 major sections: uncomplicated molecular biology, tools of gene manipulation, and sleek functions of genetic engineering. purposes coated within the e-book contain genomics, protein engineering, gene treatment, cloning, transgenic animals and crops and bioethics. An creation to Genetic Engineering is vital analyzing for undergraduate scholars of biotechnology, genetics, molecular biology, and biochemistry.
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Extra resources for An Introduction to Genetic Engineering
Fig. 4. Labelling DNA by primer extension (oligolabelling). (a) DNA is denatured to give single-stranded molecules. (b) An oligonucleotide primer is then added to give a short double-stranded region with a free 3Ј-OH group. (c) The Klenow fragment of DNA polymerase I can then synthesise a copy of the template strand from the primer, incorporating [␣-32P]dNTP (ﬁlled circles) to produce a labelled molecule with a very high speciﬁc activity. Gel electrophoresis 33 In a radiolabelling reaction it is often desirable to separate the labelled DNA from the unincorporated nucleotides present in the reaction mixture.
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. Modiﬁed from Brown (1990), Gene Cloning, Chapman and Hall; and Williams and Patient (1988), Genetic Engineering, IRL Press. Reproduced with permission.
In probe preparation the radioactive label is usually the high-energy ␤-emitter 32P. Some common methods of labelling nucleic acid molecules are described below. 1 End labelling In this technique the enzyme polynucleotide kinase is used to transfer the terminal phosphate group of ATP onto 5Ј-hydroxyl termini of nucleic acid molecules. If the ATP donor is radioactively labelled, this produces a labelled Radiolabelling of nucleic acids 31 Fig. 2. End labelling DNA using polynucleotide kinase (PNK).
An Introduction to Genetic Engineering by Dr Desmond S. T. Nicholl