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DNA-DNA hybridization of single-copy DNA sequences

Springer, Mark S. and Britten, Roy J. (1993) DNA-DNA hybridization of single-copy DNA sequences. In: Molecular Evolution: Producing the Biochemical Data. Methods in Enzymology. No.224. Academic Press , San Diego, CA, pp. 232-243. ISBN 9780121821258.

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This chapter is reviews the basic principles of hybridization and the kinetics of reassociation. It provides an overview that summarizes and compares different techniques used in single-copy hybridization. The chapter then examines different estimates of distance derived from melting curves. Native DNA is isolated and purified to remove RNA and protein. Long-stranded DNA is then sheared to short fragments to permit the separation of repetitive and single-copy DNA and to reduce viscosity and gel formation. The chapter also explains the kinetics of reassociation. Rates of reassociation of DNA are influenced by several factors including genome complexity, DNA concentration, fragment size, reassociation temperature, and cation concentration. The complexity of the sheared genome is the length in base pairs (bp) of the longest nonrepeating sequence that is produced by splicing together fragments in the population. The rate of reassociation is inversely proportional to the complexity of the genome. The hybridization of tracer DNA with driver DNA is also overviewed in the chapter.

Item Type:Book Section
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Additional Information:© 1993 Academic Press, Inc.
Series Name:Methods in Enzymology
Issue or Number:224
Record Number:CaltechAUTHORS:20160308-070622092
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Official Citation:Mark S. Springer, Roy J. Britten, [17] DNA-DNA hybridization of single-copy DNA sequences, Methods in Enzymology, Academic Press, 1993, Volume 224, Pages 232-243, ISSN 0076-6879, ISBN 9780121821258, (
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:65169
Deposited By: Ruth Sustaita
Deposited On:14 Mar 2016 21:51
Last Modified:03 Oct 2019 09:44

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