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Allele-Specific Quantification

Allele Specific Quantification

Scientific Overview

Compared to other genotyping methods and platforms, pyrosequencing technology offers the unique advantage of providing quantitative measurements of every allele. This property makes Pyrosequencing the premier choice for allele-specific expression analysis, loss of heterozygosity, and studies involving mixed populations and sample pools.

The amplification signal in Pyrosequencing is proportional to the amount of genetic material and nucleotide present in the order of the sequence. As a result, it is possible to quantify allelic variation when the amplification signals are compared to reference peaks. Ideally, their sum would equal the signal level of the reference.

epigendx_Allele Specific Quantification_chart

How It Works

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EpigenDx Allele Quantification Applications

Application: Allele-Specific Expression Analysis

Pyrosequencing technology quantifies alleles by identifying heterozygous SNPs and quantitatively measuring the amplification of each allele. This technology can help identify and mark epigenetic phenomena such as X-inactivation and genomic imprinting, situations in which multiple alleles are not expressed equally.

Application: Loss of Heterozygosity (LOH)

Loss of Heterozygosity is a biological phenomenon frequently associated with oncogenesis, usually describing a situation in which one remaining tumor-suppressor allele is silenced. This can produce various complications and malignancies. EpigenDx can detect loss of heterozygosity on a global scale using the Affymetrix CytoScan HD microarray at high resolution. Pyrosequencing technology in allele-quantification mode may detect LOH in individual groups of bases, provided enough SNP sites are covered in the analysis.

The Services Include:

Items Service Products Description Order
8024 PCR amplification PCR from genomic DNA for Pyrosequencing allele quantification analysis Inquiry / Order
8009 Allele quantification Pyrosequencing quantitative genotyping and allele quantification analysis Inquiry / Order
8006 Assay Design – Standard In silico assay design for PCR and Pyrosequencing analysis.
Customers will submit the DNA sequence containing the target region of interest. We deliver an assay design report with recommendations for PCR and Pyrosequencing primers. Customer will run PCR and Pyrosequencing based on our design.
Inquiry / Order
8007 Assay Design – Gold Assay design for Pyrosequencing analysis.
Customers submit the DNA sequence containing the target region of interest. We deliver an assay design report including PCR and Pyrosequencing primers. Customers will run PCR based on our design and send the PCR product to us for Pyrosequencing analysis.
Inquiry / Order
8008 Assay Design – Premium Assay design and assay optimization for Pyroseqencing analysis.
Customers submit the DNA sequence containing target region of interest. We optimize the PCR and Pyrosequencing assay on 10 DNA samples or controls and deliver an assay design report with PCR and Pyrosequencing primers and assay conditions along with performance data.
Inquiry / Order

Didn’t Find What You’re Looking For?

No problem, we’ve got you covered. EpigenDx provides custom assay design and analytical services with an assay database of thousands of Off-the-Shelf designs ready to go. Not only do we provide aggressive timelines with regular project updates, you will also get the highest quality technical support and consultation from design phase through project completion and data delivery. Get in touch and one of our team experts will be more than glad to assist you.

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