Journal article
Application of fluorescence-based semi-automated AFLP analysis in barley and wheat
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.
Language: | English |
---|---|
Publisher: | Springer-Verlag |
Year: | 2000 |
Pages: | 545-551 |
Journal subtitle: | International Journal of Plant Breeding Research |
ISSN: | 00405752 and 14322242 |
Types: | Journal article |
DOI: | 10.1007/s001220050071 |