Chapter 3 Manipulating DNA: From cloning to knockouts

Witkowski, Jan A. (1996) Chapter 3 Manipulating DNA: From cloning to knockouts. In: Foundations of Modern Biochemistry. JAI, pp. 27-57. ISBN 1874-5660

URL: http://www.sciencedirect.com/science/article/pii/S...
DOI: 10.1016/S1874-5660(96)80019-0

Abstract

During the 1960s, detailed genetic analysis could be performed on viruses and bacteria, but the genomes of higher organisms were too large and complex to analyze at the molecular level. Being able to isolate genes and to make copies of them by cloning was a prerequisite for the development of molecular genetics. With a gene isolated and available in large amounts, its structure and function can be analyzed directly in contrast to classical genetics that relies on inferences from observable phenotypes. Cloning is not the only way of making DNA and chemical methods had been developed long before. However, it took further developments in the early 1980s for chemical DNA synthesis to become a routine tool for molecular applications. The third method for making DNA is the polymerase chain reaction. This technique is extraordinarily versatile and has brought about a revolution in DNA and genetic manipulations. The basic strategy for cloning any fragment of DNA are (1) isolation of the DNA containing the gene of interest, (2) choosing a suitable vector-DNA capable of replicating in a host cell, (3) preparing fragments of the DNA of a size to fit into the vector, (4) introducing these recombinant DNA molecules into bacterial cells and isolate clones, and (5) identifying the clones containing the gene or DNA of interest.

Item Type: Book Section
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
Investigative techniques and equipment > cloning
Investigative techniques and equipment > assays > cloning
CSHL Authors:
Communities: Banbury Center
Depositing User: Matt Covey
Date: 1996
Date Deposited: 02 Dec 2014 20:48
Last Modified: 02 Dec 2014 20:48
URI: http://repository.cshl.edu/id/eprint/30378

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