Characterization of meiotic non-crossover molecules from Arabidopsis thaliana pollen

Meiotic recombination is essential for proper segregation of homologous chromosomes and thus for formation of viable gametes. Recombination generates either crossovers (COs), which are reciprocal exchanges between chromosome segments, or gene conversion not associated with crossovers (NCOs). Both kinds of events occur in narrow regions (less than 10 kb) called hotspots, which are distributed along chromosomes. While NCOs may represent a large fraction of meiotic recombination events in plants, as in many other higher eukaryotes, they have been poorly characterized due to the technical difficulty of detecting them. Here, we present a powerful approach, based on allele-specific PCR amplification of single molecules from pollen genomic DNA, allowing detection, quantification and characterization of NCO events arising at low frequencies at recombination hotspots.