Visualization of Differential Gene Expression– Using Fluorescence-Based cDNA-AFLP

cDNA-AFLP is one of the techniques developed to study differentially expressed genes. Ibis recent technique is advantageous because it does not need prior sequence knowledge and is reliable due to highly stringent PCR conditions. The traditional cDNA-AFLP method uses radioactively labelled products and is characterised by high sensitivity and resolution. Here, the use of Cy5-labelled primers to detect products on polyacrylamide gels is reported. This non-radioactive method, based on fluorescence, is shown to be faster and the recovery of interesting bands is easier. The study of the differential gene expression of the interaction between potato and Phytophthora infestans was used for the valuation of this method. Different gene expression profiles - such as up-regulation, down-regulation or point expression - were obtained. Moreover, this technique was shown to be highly reproducible.