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therascreen UGT1A1 Pyro Kit

For genotyping of the allele variants *28 and *6 of the human UGT1A1 gene
  • Compliance with EU IVD Directive 98/79/EC
  • Results in real time
  • Easy interpretation of complex sequence information
  • Comprehensive post-run analysis
The therascreen 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.
Cat No./ID: 971540
therascreen UGT1A1 Pyro Kit (24)
For 24 reactions: Sequencing Primers, PCR Primers, Human Control DNA, PyroMark PCR Master Mix, CoralLoad Concentrate, Buffers, and Reagents

The therascreen UGT1A1 Pyro Kit is intended for in vitro diagnostic use.


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Pyrogram trace obtained after analysis of a sample with –/TA (TA6/TA7) genotype when analyzed for allele variant *28.

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Pyrogram trace obtained after analysis of samples with a G/G genotype when analyzed for allele variant *6.

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Pyrogram trace obtained after analysis of a sample with –/– (TA6/TA6) genotype when analyzed for allele variant *28.

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Illustrations of the therascreen 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 therascreen 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. The UGT1A1 gene encodes the enzyme UDP-glucuronosyltransferase, which catalyzes the conversion of xenobiotic and endogenous compounds into water-soluble molecules that can be excreted. Genetic polymorphisms found in the human UGT1A1 gene are associated with reduced UGT1A1 expression and activity. The most common polymorphisms occur in the UGT1A1 promoter region that contains several 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 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.
Applications
The therascreen UGT1A1 Pyro Kit for genotyping of allele variant *28 and allele variant *6 of the human UGT1A1 gene (see figures "Pyrogram trace obtained after analysis of a sample with –/– (TA6/TA6) genotype when analyzed for allele variant *28", "Pyrogram trace obtained after analysis of a sample with –/TA (TA6/TA7) genotype when analyzed for allele variant *28", and "Pyrogram trace obtained after analysis of samples with a G/G genotype when analyzed for allele variant *6"). This kit enables genotyping analysis applicable to several fields of clinical applications, such as diagnostic evaluation, clinical research, and pharma/biopharma testing.
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