The kit consists of two assays: one for genotyping of allele variant *28 and the second for genotyping of allele variant *6. The two regions are amplified separately by PCR and sequenced through the defined region (see figure " Illustrations of the therascreen UGT1A1 assays
"). Sequences surrounding the defined positions serve as normalization and reference peaks for genotyping and quality assessment of the analysis.
After PCR using primers targeting allele variants *28 and *6, the amplicons are immobilized on Streptavidin Sepharose High Performance beads. Single-stranded DNA is prepared, and the corresponding sequencing primers anneal to the DNA. The samples are then analyzed on the PyroMark Q24 system.