cDNA-AFLP-based transcript profiling for genome-wide expression analysis of jasmonate-treated plants and plant cultures

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1011; p. 287
• Main Authors: Colling, Janine, Pollier, Jacob, Makunga, Nokwanda P, Goossens, Alain
• Format: Journal Article
• Language: English
• Published: United States 2013
• Subjects: Amplified Fragment Length Polymorphism Analysis; Culture Techniques; Cyclopentanes - pharmacology; DNA, Complementary - genetics; Gene Expression; Gene Expression Profiling; Genome, Plant; Oxylipins - pharmacology; Plant Growth Regulators - pharmacology; Plant Proteins - genetics; Plant Proteins - metabolism; Polymerase Chain Reaction
• ISSN: 1940-6029
• DOI: 10.1007/978-1-62703-414-2_23

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.