This is an example for real time monitoring of the dye-labeled oligonucleotide ligation (DOL) assay. The assay consists of two consecutive reactions, a PCR reaction and a ligation reaction. The fluorescence intensities for the donor (FAM, right axis) and the acceptors (ROX and TAMRA, left axis) stay unchanged in the PCR reaction, once the ligation reaction starts, the ROX intensity is steadily going up in corresponding to the decreasing of FAM intensity. This is a positive result for ROX labeled allele. In contrast, TAMRA intensity did not show the trend of increasing, so the TAMRA labeled allele is negative. For a heterozygote, both ROX and TAMRA intensities will increase.

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Last revised: January 13, 2004.