Plant & Animal Genomes XIII Conference

January 15-19, 2005
Town & Country Convention Center
San Diego, CA

GeneMining(tm) Differential Expression Kit, A Simple And Effective Approach In Gene Discovery

Fan Zhang , Keming Song

  Sigma-Aldrich Corporation, 2909 Laclede Ave., St. Louis, MO 63103

We have developed a GeneMiningTM Differential Expression system as a simple and effective tool for gene discovery and differential expression profiling. The system is a RT-PCR based differential display method using a proprietary ACP (Annealing Control Primer) technology. The ACP technology features a set of specially designed primers that comprise a tripartite structure with a polydeoxyinosine [poly(dI)] linker between the 3’end target core sequence and the 5’ end non-target universal sequence. This unique structure prevents annealing of the 5’ end non-target sequence to the template and thus facilitates primer hybridization at the 3’ end to the target sequence at specific temperatures, resulting in a dramatic improvement of annealing specificity. Compared to existing differential display methods, the GeneMiningTM Differential Expression system provides superior reproducibility and sensitivity. The amplicons produced by this new method can be displayed on regular agarose gel, making it easy for band excision, purification, and sequencing. Also, the whole process can be carried out in a high throughput fashion. We have conducted extensive validation of the ACP technology in Arabidopsis, corn, soybean, and mammalian systems (CHO and HeLa cells). Comparison between our data with the data from microarray and/or real time RT-PCR indicated that GeneMiningTM Differential Expression system is a simple, reliable, and cost-effective tool for gene discovery and expression profiling, particularly useful for plant and animal species where genome sequence information is often limited.