Abstract
We are interested in cloning/identifying virtually all mammalian brain mRNAs which are differentially expressed in response to various chemical, electrical, pharmacological and behavioral challenges to the central nervous system. To reach this ambitious objective, we propose that a PCR-based differential display methodology which is digital, portable, systematic, quantitative and database-friendly is required. To develop this technology, we first examined previously reported PCR-based differential display methodologies; we report that although current methods are sufficient for identifying some differentially expressed mRNAs, a more rigorous technique is needed to fit the necessary criteria. To this end, we report preliminary work which is aimed at the development of a modified method of PCR-based differential display.
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References
M.B. Dworkin and I.B. Dawid, Construction of a cloned library of expressed embryonic gene sequences from Xenopus laevis. Dev. Biol. 76:435 (1980).
J.D. Falk, H. Usui, and J.G. Sutcliffe, Identification of expressed sequences on human chromosome 9q32–34, Proceed. Transcribed Seq. Workshop,in press (1994).
H. Usui, J. Falk, A. Dopazo, L. de Lecea, M.G. Erlander, and J.G. Sutcliffe, Isolation of clones of rat striatum-specific mRNAs by directional tag PCR subtraction, J. Neurosci. In Press, (1994).
J.G.K. Williams, A.R. Kubelik, K.J. Livak, J.A. Rafalski, and S.V. Tingey, DNA polymorphisms amplified by arbitrary primers are useful as genetic markers, Nucl. Acid Res. 18:6531 (1990).
J. Welsh and M. McClelland, Fingerprinting genomes using PCR with arbitrary primers, Nucl. Acids Res. 18:7213 (1990).
J. Welsh and M. McClelland, Genomic fingerprinting using arbitrarily primed PCR and a matrix of pairwise combinations of primers, Nucl. Acids Res. 19:5275 (1991).
S.R. Woodward, J. Sudweeks, and C. Teuscher, Random sequence oligonucleotide primers detect polymorphic DNA products which segregate in inbred strains of mice, Mammal. Genome 3:73 (1992).
J.H. Nadeau, H.G. Bedigian, G. Bouchard, T. Denial, M. Kosowsky, R. Norberg, S. Pugh, E. Sargeant, R. Turner, and B. Paigen, Multilocus markers for mouse genome analysis: PCR amplification based on single primers of arbitrary nucleotide sequence, Mammal. Genome 3:55 (1992).
P. Liang, and A.B. Pardee, Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction, Science 257:967 (1992).
J. Welsh, K. Chada, S.S. Dalai, R. Cheng, D. Ralph, and M. McClelland, Arbitrarily primed PCR fingerprinting of RNA, Nucl. Acids Res. 20:4965 (1992).
P. Liang, L. Averboukh, and A.B. Pardee, Distribution and cloning of eukaryotic mRNAs by means of differential display: refinements and optimization, Nucl. Acids Res. 21:3269 (1993).
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© 1994 Springer Science+Business Media New York
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Erlander, M.G., Dopazo, A., Foye, P.E., Sutcliffe, J.G. (1994). PCR-Based Technologies to Study Differential Gene Expression in Rat Brain. In: Hochgeschwender, U., Gardiner, K. (eds) Identification of Transcribed Sequences. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-2562-2_23
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DOI: https://doi.org/10.1007/978-1-4615-2562-2_23
Publisher Name: Springer, Boston, MA
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