For genotyping of the allele variant *28 and allele variant *6 of the human UGT1A1 gene
  • 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.

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UGT1A1 Pyro Kit (24)
For 24 reactions: Sequencing Primers, PCR Primers, Control DNA, PyroMark PCR Master Mix, CoralLoad Concentrate, Buffers, and Reagents
970540
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The UGT1A1 Pyro Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Pyrogram trace obtained after analysis of a sample with -/- (TA6/TA6) genotype when analyzed for allele variant *28|Pyrogram of sample with -/TA (TA6/TA7) genotype, analyzed for allele variant *28.|Pyrogram trace obtained after analysis of samples with a G/G genotype when analyzed for allele variant *6|Illustrations of the UGT1A1 assays|
|||The sequence indicated is the analyzed sequence with polymorphic nucleotides indicated by square brackets or slash. Part of the TA repeats analyzed with the UGT1A1 *28 assay are covered by the sequencing primer. FP, FPB: Forward PCR primers (B indicates biotinylation); RP, RPB: Reverse PCR primers (B indicates biotinylation); Seq: Sequencing primers.|
Principle

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.

Procedure

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").

Applications

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.

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Kit Handbooks
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Safety Data Sheets
1