Schematic illustration of MOL-PCR.

<p>A. Three oligonucleotides and two fluorescently labelled beads are used for interrogation of one SNP. B. MOL-PCR consists of three steps. 1. One of two competitive allele-specific left-hand probe oligonucleotides (LPO) and one universal right-hand probe oligonucleotide (RPO) are hybridized to the template DNA and ligated. 2. With PCR and a reporter-labelled forward primer, the ligated oligonucleotides (LPO+RPO) are amplified. 3. After denaturation of the PCR product, allele-specific fluorescent beads carrying an allele-specific <i>antiTag</i> sequence are hybridized to the amplicons. This will result in beads carrying reporter fluorescence (bead 1 in example) and beads not carrying fluorescence (bead 2). Reporter fluorescence (Alexa532) per bead is measured with a flow cytometric device (Luminex). For the 8-plex assay, a total of 24 oligonucleotides and 16 beads are used in the same reaction.