Abstract
Nucleic acid probes are an important tool in molecular diagnosis. To facilitate the detection of hybridized probes, they are labeled with a reporter molecule, which is usually a radioisotope. For diagnostic techniques carried out in a clinical laboratory, radioisotopes are hazardous and, thus, recently there is a move to use nonisotopic labels, such as biotin and digoxigenin. Nonisotopic probes also have the advantage of much better stability over time compared with isotopic probes that have limited half-lives.
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
References
Rigby, P. W. J., Dieckmann, M., Rhodes, C., and Berg, P. (1977) Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase. J. Mol. Biol. 113, 237–247.
Feinberg, A. P. and Vogelstein, B. (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132, 6–13.
Lo, Y. M. D., Mehal, W. Z., and Fleming, K. A. (1988) Rapid production of vector-free biotinylated probes using the polymerase chain reaction. Nucleic Acids Res. 16, 8719.
An, S. F., Franklin, D., and Fleming, K. A. (1992) Generation of digoxigenin-labeled double-stranded and single-stranded probes using the polymerase chain reaction. Mol Cell. Probes 6, 193–200.
Chan, V. T. W., Fleming, K. A., and McGee, J. O. (1985) Detection of subpicogram quantities of specific DNA sequences on blot hybridisation with biotinylated probes. Nucleic Acids Res. 13, 8083–8091.
Boehringer Mannheim. (1989) Biochemica-Applications Manual: DNA Labelling and Nonradioactive Detection. Boehringer Mannheim GmbH Biochemica, East Sussex, UK.
Barnes, W. M. (1994) PCR amplification of up to 35 kb DNA with high fidelity and high yield from Lambda bacteriophage templates. Proc. Natl. Acad. Sci. USA 91, 2216–2220.
Weir, H. U. G., Segraves, R., Pinkel, D., and Gray, J. W. (1990) Synthesis of Y chromosome-specific labelled DNA probes by in vitro DNA amplification. J. Histochem. Cytochem. 38, 421–426.
Duplaa, C., Couffinhal, T., Labat, L., Moreau, C., Lamaziere, J., and Bonnet, J. (1993) Quantitative analysis of polymerase chain reaction products using biotinylated dUTP incorporation. Anal. Biochem. 212, 229–236.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2000 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Dennis Lo, Y.M., An, S.F. (2000). Generation of Labeled Probes by Polymerase Chain Reaction. In: Rapley, R. (eds) The Nucleic Acid Protocols Handbook. Springer Protocols Handbooks. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-038-1:127
Download citation
DOI: https://doi.org/10.1385/1-59259-038-1:127
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-459-4
Online ISBN: 978-1-59259-038-4
eBook Packages: Springer Book Archive