Cloning, DNA

The principle of Consider an experimental goal which is to make large amounts of a particular DNA cloning DNA fragment in pure form from a mixture of DNA fragments. Although the 

DNA fragments can be introduced into bacterial cells, most or all will lack the ability for self-replication and will quickly be lost. However, two types of DNA molecule are known which can replicate autonomously in bacterial cells bacteriophages (see Topic 14) and plasmids. Plasmids are small circular double-stranded DNA molecules that exist free inside bacterial cells, often carry 

Instant Notes in Biochemistry 2nd Edition, B.D. Hames N.M. Hooper, (c) 2000 BIOS Scientific Publishers Ltd, Oxford. 

A DNA library is a collection of cloned DNA fragments in a cloning vector that can be searched for a DNA of interest. If the goal is to isolate particular gene sequences, two types of library are useful  

Each bacterial colony is a separate clone of bacterial cells each containing the same recombinant plasmid 

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Chapter 13 Recombinant DNA Cloning and Creation of Chimeric Genes... 

Rebmann G. Hertig C. Bull J. Mauch F. Dudler R. (1991) Complementary DNA cloning and sequence analysis of a pathogen-induced putative peroxidase from rice / / Plant Mol. Biol. V. 16. P. 329-331. 

DNA cloning and molecular biology coupled with protein chemistry provide a hybrid approach that greatly increases the speed and efficiency for determination of primary strucmres of proteins. 

In DNA cloning, a particular segment of DNA is removed from its normal environment using one of many restriction endonucleases. This is then ligated into one of several vectors in which the DNA segment can be ampfified and produced in abundance. 

CHARACTERIZATION OF GREEN BEAN PE (C)DNA CLONES 4.1 Isolation of (c)DNA clones... 

Hartley, J. L., Temple, G. F., and Brasch, M. A. (2000). DNA cloning using in vitro site-specific recombination. Genome Research 10, 1788-1795. 

Gorman, C. (1985). High efficiency gene transfer into mammalian cells. In Glover, D. (Ed) DNA cloning II. A Practical Approach, pp. 143—190. IRL Press Ltd., Oxford, UK. 

Figure 3.12 A basic overview of the DNA cloning process. Refer to text for specific details...

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