Abstract
The polymerase chain reaction (PCR) is an incredibly versatile technique that has made a profound impact on many areas of biology. PCR is based on the use of oligonucleotides, which flank the region of DNA of interest and which are complementary to sequences on either DNA strand, to prime the replication of each strand by Taq DNA polymerase. This enzyme is thermostable, and consequently, one can go through cycles of template denaturation and renaturation after each round of replication and exponentionally amplify the sequence of interest. This procedure is very sensitive, and as such, it allows one to amplify, isolate, and utilize specific DNA sequences from vanishingly small quantities of starting material using either genomic or cDNA as a substrate.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Sambrook, J., Fritsch, E. F., and Maniatis, T. (eds.) (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Chomczynski, P. and Sacchi, N. (1987) Single step method of RNA isolation by guanindinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162, 156–159.
Childs, S. R., Wrana, J. L., Arora, K., Attisano, L., O’Connor, M. B., and Massague, J. (1993) Identification of a Drosophila activin receptor. Proc. Natl. Acad. Sci. USA 90, 9475–9479.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Humana Press Inc.
About this protocol
Cite this protocol
Graham, A. (1999). RT-PCR on Embryos Using Degenerate Oligonucleotide Primers. In: Sharpe, P.T., Mason, I. (eds) Molecular Embryology. Methods in Molecular Biology™, vol 97. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-270-8:591
Download citation
DOI: https://doi.org/10.1385/1-59259-270-8:591
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-387-0
Online ISBN: 978-1-59259-270-8
eBook Packages: Springer Protocols