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

G. Schwarz, M. Herz, X.Q. Huang, W. Michalek, A. Jahoor, G. Wenzel, V. Mohler

    Research output: Contribution to journalJournal articleResearch

    Abstract

    Genetic mapping and the selection of closely linked molecular markers for important agronomic traits require efficient, large-scale genotyping methods. A semi-automated multifluorophore technique was applied for genotyping AFLP marker loci in barley and wheat. In comparison to conventional P-33-based AFLP analysis the technique showed a higher resolution of amplicons, thus increasing the number of distinguishable fragments. Automated sizing of the same fragment in different lanes or different gels showed high conformity, allowing subsequent unambigous allele-typing. Simultaneous electrophoresis of different AFLP samples in one lane (multi-mixing), as well as simultaneous amplification of AFLP fragments with different primer combinations in one reaction (multiplexing), displayed consistent results with respect to fragment number, polymorphic peaks and correct size-calling. The accuracy of semi-automated codominant analysis for hemizygous AFLP markers in an F-2 population was too low, proposing the use of dominant allele-typing defaults. Nevertheless, the efficiency of genetic mapping, especially of complex plant genomes, will be accelerated by combining the presented genotyping procedures.
    Original languageEnglish
    JournalTheoretical and Applied Genetics
    Volume100
    Issue number3-4
    Pages (from-to)545-551
    ISSN0040-5752
    DOIs
    Publication statusPublished - 2000

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