Open AccessVisualization of Differential Gene Expression – Using Fluorescence‐Based cDNA‐AFLP
Author(s) -
Gigliotti S.,
Hausman J.F.,
Evers D.
Publication year - 2004
Publication title -
engineering in life sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.547
H-Index - 57
eISSN - 1618-2863
pISSN - 1618-0240
DOI - 10.1002/elsc.200420002
Subject(s) - complementary dna , biology , gene , gene expression , amplified fragment length polymorphism , phytophthora infestans , microbiology and biotechnology , differential display , computational biology , genetics , population , demography , sociology , genetic diversity
cDNA‐AFLP is one of the techniques developed to study differentially expressed genes. This 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.