|Values are valid only on day of printing.|
Genomic DNA is extracted from whole blood.
Luminex Assay (tier 1): The xTAG assay for CYP2D6 incorporates multiplex PCR and multiplex allele-specific primer extension (ASPE) with Luminex Molecular Diagnostics' proprietary universal tag sorting system on the Luminex 100 xMAP platform.(Package insert: xTAG Kit for CYP2D6, Luminex Molecular Diagnostics Inc, Austin, TX)
Copy Number Assay (tier 2): This assay utilizes a duplex real-time PCR, which includes 1 copy number probe (FAM labeled) and a reference assay (VIC labeled) per reaction. Each copy number probe detects the genomic sequence of interest and the reference assay detects a sequence that is known to be present in 2 copies in a diploid genome. Relative quantitation is used to determine the relative copy number of the target of interest in a genomic DNA (gDNA) sample normalized to 10 ng/mcL for each probe. Each probe is normalized to the known copy number of the reference sequence, and compared to a calibrator sample with known copies of the target sequence included with each run.(Package insert: Taqman Copy Number Assays Revision D, Applied Biosystems, Carlsbad, CA)
Sequencing component assays (tier 3): The CYP2D6 gene of interest is amplified by PCR. The PCR product is then purified and sequenced in both directions using fluorescent dye-terminator chemistry. Sequencing products are separated on an automated sequencer and trace files analyzed for variations in the exons and intron/exon boundaries of all 9 exons using mutation detection software and visual inspection.(Unpublished Mayo method)
Monday, Thursday; 8 a.m.