Application of fluorescence based semi-automated AFLP analysis in barley and wheat

Genetic mapping and the selection of closely linked molecular markers for i mportant agronomic traits require efficient, large-scale genotyping methods . A semi-automated multifluorophore technique was applied for genotyping AF LP marker loci in barley and wheat. In comparison to conventional P-33-base d AFLP analysis the technique showed a higher resolution of amplicons, thus increasing the number of distinguishable fragments. Automated sizing of th e same fragment in different lanes or different gels showed high conformity , allowing subsequent unambigous allele-typing. Simultaneous electrophoresi s of different AFLP samples in one lane (multi-mixing), as well as simultan eous amplification of AFLP fragments with different primer combinations in one reaction (multiplexing), displayed consistent results with respect to f ragment number, polymorphic peaks and correct size-calling. The accuracy of semi-automated codominant analysis for hemizygous AFLP markers in an F-2 p opulation was too low, proposing the use of dominant allele-typing defaults . Nevertheless, the efficiency of genetic mapping, especially of complex pl ant genomes, will be accelerated by combining the presented genotyping proc edures.