UGT1A1 Pyro Kit

• Comprehensive results in real time
• Easy interpretation of complex sequence information
• Sequence context as built-in control of the assay
• Flexible post-run analysis

The UGT1A1 Pyro Kit is used for genotyping of allele variant *28 and allele variant *6 of the human UGT1A1 gene. The kit contains primers and reagents for amplification of the genes, plus buffers, primers, and reagents for genotyping in real time using Pyrosequencing technology on the PyroMark Q24 System.

The UGT1A1 Pyro Kit is used for genotyping of allele variant *28 (for distinguishing between 6 and 7 TA repeats) and allele variant *6 (for distinguishing between G and A genotype) of the human UGT1A1 gene in real time using Pyrosequencing technology on the PyroMark Q24 System. The UGT1A1 gene encodes the enzyme UDP-glucuronosyltransferase that catalyzes the conversion of compounds into water-soluble molecules to be excreted. Genetic polymorphisms found in the UGT1A1 gene are associated with reduced expression and activity. Polymorphisms are most commonly found in the UGT1A1 promoter region that contains repeating TA elements (UGT1A1*28) and as a non-synonymous coding variant G71R (UGT1A1*6) in exon 1.

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 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 (see " Pyrogram trace of a -/- (TA6/TA6) genotype when analyzed for allele variant *28", " Pyrogram trace of -/TA (TA6/TA7) genotype when analyzed for allele variant *28", and " Pyrogram trace of a G/G genotype when analyzed for allele variant *6").

The UGT1A1 Pyro Kit is intended for genotyping of allele variant *28 (for distinguishing between 6 and 7 TA repeats) and allele variant *6 (for distinguishing between G and A genotype) of the human UGT1A1 gene.