Plant & Animal Genome VI Conference

P80

COMPARISON OF ³²P, AUTOMATED FLORESCENCE, AND SILVER STAINING VISUALIZATION METHODS FOR DETECTING AFLPS

1. Washington State University, USDA-ARS Wheat Genetics, 209 Johnson Hall , Pullman, WA 99164-6420
2. Washington State University, Dept. of Crop and Soil Sciences, 201 Johnson Hall, Pullman, WA 99164-6420

The utility of a DNA marker system is determined by the consistency and efficiency of the detection method used. Amplified fragment length polymorphism (AFLP) technology is an effective marker system ammenable to several visualization techniques, and is readily automated. The objective of this research was to determine whether ³²P, automated florescence, and silver staining detection methods generate uniform, reproducable AFLP data. Using genomic DNA from hexaploid wheat (Triticum aestivum L.) cultivars, AFLP analyses were conducted with restiction enzyme combinations PstI:MseI or EcoRI:MseI, using PCR conditions optimized for each detection method. Fragment data obtained from digitized images of autoradiographs and silver stained gels were compared to each other using ProScore^TM software, and those images also were compared to images from the ABI PRISM^TM florescence-based Automated Genotyping System^TM. Quantitative and qualitative measures of the consistency with which each system detected polymorphisms were determined. Results will be used to ease transfer of informative AFLP markers among wheat research groups.