UGT1A1 Pyro Kit

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

Features

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

Useful Resources

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UGT1A1 Pyro Kit (24)

Cat. No. / ID: 970540

For 24 reactions: Sequencing Primers, PCR Primers, Control DNA, PyroMark PCR Master Mix, CoralLoad Concentrate, Buffers, and Reagents
The UGT1A1 Pyro Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

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.

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").

See figures

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.

Supporting data and figures

Resources

Kit Handbooks (1)
Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.

FAQ

Can I use more than 10ng DNA for the PCR reaction for the Pyro kits?
The NRAS Pyro, EGFR Pyro. BRAF PyroKRAS Pyro, and UGT1A1 Pyro Kits all require that no more than 10ng of DNA be used for the PCR reaction.  Too much DNA can sometimes lead to inhibition problems in the PCR reaction.
FAQ ID -2379
Can PyroMark Gold reagents be vortexed?
Reconstiuted enzyme and substrate of PyroMark Gold Reagents, should not be vortexed since this could lead to conformational changes which affect the activity.
FAQ ID -2844