Abstract
cDNA-AFLP is a commonly used, robust, and reproducible tool for genome-wide expression analysis in any species, without requirement of prior sequence knowledge. Quantitative expression data are generated by gel-based visualization of cDNA-AFLP fingerprints obtained by selective PCR amplification of subsets of restriction fragments from a double-stranded cDNA template. Differences in gene expression levels across the samples are reflected in different band intensities on the high-resolution polyacrylamide gels. The differentially expressed genes can be identified by direct sequencing of re-amplified cDNA-AFLP tags purified from the gels. The cDNA-AFLP technique is especially useful for profiling of transcriptional responses of jasmonate-treated plants or plant (tissue) cultures and the discovery of jasmonate-responsive genes.
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Acknowledgments
This work was supported by the European Framework Programme 7 project SMARTCELL (FP7-KBBE-222716). J.C. is indebted to the Ghent University for a “Bijzondere Onderzoeksfonds” predoctoral fellowship.
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Colling, J., Pollier, J., Makunga, N.P., Goossens, A. (2013). cDNA-AFLP-Based Transcript Profiling for Genome-Wide Expression Analysis of Jasmonate-Treated Plants and Plant Cultures. In: Goossens, A., Pauwels, L. (eds) Jasmonate Signaling. Methods in Molecular Biology, vol 1011. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-414-2_23
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DOI: https://doi.org/10.1007/978-1-62703-414-2_23
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