Abstract
Nyagaard and Hall, and Southern (Nyagaard and Hall, 1964 Southern, 1975) introduced immobilizing target DNA to membrane for hybridization studies. The technique permits hybridization of various probes to immobilized target DNA under controlled conditions, coupled with fast detection of the hybrid. Because of the sensitivity, speed, and convenience of this “solid state” hybridization procedure, it has been widely applied in applied and basic research. The Southern hybridization procedure described in this chapter is separated into two general protocols: transfer of the DNA to the membrane and hybridization of immobilized DNA to DNA or RNA probes. A description of agarose gel electrophoresis of DNA is given in Chapter 8.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Benton WD, Davis RW (1977) Screening ?,,gt recombinant clones by hybridization to single plaques in situ. Science 196:180-182.
Chomczynski P (1992) One-hour downward alkaline capillary transfer for blotting of DNA and RNA. 201:134-139.
Grunstein M, Hogness D (1975) Colony hybridization: A method for isolation of cloned DNAs that contain a specific gene. Proc Nail Acad Sci USA 72:3961-3965.
Genius system user's guide for membrane hybridization. Version 3. (1995) BoehringerMannheim Biochemicals. 9115. Hague Road. P.O. Box 50414. Indianapolis, IN 46250. Hanahan D, Meselson M (1980) A protocol for high density screening of plasmids in1776. Gene 10:63-68.
Nyagaard AP, Hall BD (1964) J Mol Biol 9:125-130.
Peferoen M, Huybrecht R, De Loof A 1982 Vacuum-blotting: A new simple and efficient transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to nitrocellulose. FEBS Lett 145:369-372.
Smith MR, Devine CS, Cohn SM, Lieberman MW 1984 Quantitative electrophoretic transfer of DNA from polyacrylamide or agarose gels to nitrocellulose. Anal Biochem 137:120-124.
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:502-517.
Wei Y-G, Surzycki SJ (1986) Screening recombinant clones containing sequences homologous to Escherichia coli genes using single-stranded bacteriophage vector. Gene 48:251-256
Rights and permissions
Copyright information
© 2000 Springer-Verlag Berlin Heidelberg
About this chapter
Cite this chapter
Surzycki, S. (2000). DNA Transfer and Hybridization. In: Basic Techniques in Molecular Biology. Springer Lab Manuals. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-56968-5_11
Download citation
DOI: https://doi.org/10.1007/978-3-642-56968-5_11
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-540-66678-3
Online ISBN: 978-3-642-56968-5
eBook Packages: Springer Book Archive