Characterization of Clones
The purpose of this chapter is to introduce the plant researcher with little or no experience of DNA sequencing to a technology central to many molecular biology projects. DNA sequencing technology continues to develop at a rapid rate, and refinements to methods are reported almost daily, largely in response to the demands of ambitious projects aiming to sequence the complete genome of organisms as diverse as man, yeast (Saccharomyces cerevisiae), plants (Arabidopsis) and nematodes (Caenorhabditis elegans). It is beyond the scope of this chapter to provide a comprehensive review of every new development, but by using the protocols provided, good-quality sequence should be obtained routinely. An increasing number of laboratories have access to automated sequencing facilities. However, for many small laboratories, and those in which molecular biology is a new venture, manual methods of DNA sequencing will continue to be required. It is at plant molecular biologists working in these laboratories that this chapter is aimed.
KeywordsAmmonium Persulphate Mung Bean Nuclease Correct Reading Frame Nest Deletion Thermostable Polymerase
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