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qBiomarker Somatic Mutation PCR Arrays

For rapid and accurate profiling of the somatic DNA mutation status of gene panels

  • Pathway- or disease-focused profiling of somatic mutation status
  • Simple real-time PCR procedure
  • High sensitivity and wide dynamic range
  • Designed for routine use on most PCR instruments
  • Master mix included

qBiomarker Somatic Mutation PCR Arrays are translational research tools that allow rapid and accurate profiling of the somatic mutation status for important genes related to a biological pathway or disease. Mutations are selected from comprehensive somatic mutation databases (e.g., COSMIC) and peer-reviewed scientific literature based on their clinical or functional relevance and frequency of occurrence.

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qBiomarker Somatic Mutation PCR Arrays are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


Performance

qBiomarker Somatic Mutation PCR Arrays show a high degree of sensitivity even with formalin-fixed paraffin-embedded (FFPE) samples (see figures "High degree of sensitivity" and "Sensitivity of qBiomarker Somatic Mutation PCR Arrays with FFPE samples").

qBiomarker Somatic Mutation PCR Arrays have utility in the detection of mutations in cell lines or research samples, which is critical for toxicological, drug development, and cancer studies (see figure "Profiling of common cancer cell lines for somatic mutation status").

Principle
Real-time PCR is the most sensitive and reliable method for the detection of DNA mutations. By combining allele-specific amplification and hydrolysis probe detection, qBiomarker Somatic Mutation real-time PCR assays have been developed which can detect as few as 1% somatic mutations in the background of wild-type genomic DNA. Allele-specific amplification is achieved by Amplification Refractory Mutation System (ARMS) technology, which is based on the discrimination by Taq polymerase between a match and a mismatch at the 3’ end of the PCR primer (see figure "Principle of ARMS technology").
Procedure

Simply mix the DNA template with the ready-to-use PCR mastermix, aliquot equal volumes to each well of the same plate, and then run the real-time PCR cycling program. qBiomarker Somatic Mutation PCR Arrays are compatible with all ABI, Bio-Rad, Eppendorf, Roche, and Stratagene instruments.

qBiomarker Somatic Mutation PCR Arrays are available in 96-well and 384-well plates and are used to detect mutations related to a disease state or pathway, plus gene copy number controls for normalization. Each qBiomarker Somatic Mutation PCR Array also includes control elements for general PCR performance.

Easy-to-use data analysis

Data can be analyzed using the available Excel-based data analysis templates. Data analysis is based on either the ∆∆CT or Average ∆CT method.

Arrays are available in a variety of formats, all with mastermix included:

qBiomarker Somatic Mutation PCR Array Format A: Fluoroscein, 96-well; for Bio-Rad iCycler, iQ5, MyiQ, and MyiQ2 instruments
qBiomarker Somatic Mutation PCR Array Format A: ROX, 96-well; for ABI Standard 96-well Blocks (5700, 7000, 7300, 7500, 7900HT, ViiA 7); Bio-Rad Chromo 4 (MJ Research); Stratagene Mx3005p, Mx3000p; Eppendorf ep realplex 2/2S, and 4/4S instruments
qBiomarker Somatic Mutation PCR Array Format C: ROX, 96-well; for ABI 7500 FAST 96-well Block, 7900HT FAST 96-Well Block, StepOnePlus, and ViiA 7 FAST 96-well Block instruments
qBiomarker Somatic Mutation PCR Array Format D: ROX, 96-well; for Bio-Rad CFX96, Opticon and Opticon 2 (MJ Research); Stratagene Mx4000 instruments
qBiomarker Somatic Mutation PCR Array Format E: ROX, 384-well; for ABI 7900HT 384-well Block, ViiA 7 384-well Block; Bio-Rad CFX384 instruments
qBiomarker Somatic Mutation PCR Array Format F: ROX, 96-well; for Roche LightCycler 480 96-well Block instruments
qBiomarker Somatic Mutation PCR Array Format G : ROX, 384-well; for Roche LightCycler 480 96-well Block instruments
Applications

qBiomarker Somatic Mutation PCR Arrays are highly suited for the rapid and accurate profiling of mutations for a pathway- or disease-focused set of genes and key downstream and associated signaling genes.

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Instrument Technical Documents (2)
For screening disease-focused mutation panels by PCR
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For gene expression and genomic analysis
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Kit Handbooks (1)
For real-time PCR-based, pathway- or disease-focused somatic mutation profiling
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Download Files (1)
Data analysis file for qBiomarker™ Somatic Mutation PCR Array Human DNA QC Pathway- FFPE Samples
Catalog number- 337021
Pathway number- SMH-999

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References
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Brain Cancer qBiomarker Somatic Mutation PCR Array
Introduction
The Human Brain (GBM/CNS) Cancer qBiomarker Somatic Mutation PCR Array is a translational research tool that allows rapid, accurate, and comprehensive profiling of the top somatic mutations in human GBM/CNS cancer in the following genes: BRAF, CTNNB1/beta-catenin, EGFR, IDH1, IDH2, KRAS, NF2, NRAS, PIK3CA, and PTEN. These mutations warrant extensive investigation to enhance the understanding of carcinogenesis and identify potential drug targets. Numerous research studies have demonstrated the utility of individual and multiple somatic mutation status information in identifying key signaling transduction disruptions. For example, the mutation status of the EGFR and KRAS genes can predict the physiological response to certain drugs targeting these molecules. The Human GBM/CNS Cancer qBiomarker Somatic Mutation PCR Array, with its comprehensive content coverage, is designed for the study of mutations in the context of GBM/CNS cancers and has the potential for discovery and verification of drug target biomarkers for these cancer types and other cancer types in which these mutations have been identified. This array includes 37 DNA sequence mutation assays designed to detect the most frequent, functionally verified, and biologically significant mutations in human GBM/CNS cancers. These mutations were chosen from curated, comprehensive somatic mutation databases and peer-reviewed scientific literature, and represent the most frequently recurring somatic mutations compiled from over 7800 CNS cancer samples (including 6850 glioma samples). Each 96-well array allows profiling of the mutation status of 2 samples, while each 384-well format array allows mutation profiling of 8 samples. The simplicity of the product format and operating procedure allows routine somatic mutation profiling in any research laboratory with access to real-time PCR instruments.
Gene List

BRAF: 1 Assay
The most important BRAF mutation in brain cancer leads to increased kinase activity, the p. V600E mutation.

CTNNB1: 5 Assays
The most frequently detected CTNNB1/beta-catenin mutations result in abnormal signaling in the WNT signaling pathway. The mutated codons are mainly several serine/threonine residues targeted for phosphorylation by GSK-3beta.

EGFR: 4 Assays
The most frequently identified EGFR mutations include P-loop and activation loop point mutations, kinase domain deletions, and insertion mutations.

IDH1: 5 Assays
Most of these mutations abolish magnesium binding and alters the enzyme's activity to convert alpha-ketoglutarate into R(-)-2-hydroxyglutarate instead of isocitrate into alpha-ketoglutarate.

IDH2: 3 Assays
These mutations all lie in the substrate binding domain, and one (p.R140Q) is associated with D-2-hydroxyglutaric aciduria.

KRAS: 2 Assays
The mutation assays include the most frequently occurring mutations in KRAS codons 12, 13, and 61. Mutations at these positions result in reduced intrinsic GTPase activity and/or cause KRAS to become unresponsive to RasGAP.

NF2: 1 Assay
NF2 is similar to some members of the ERM (ezrin, radixin, moesin) family of proteins and links cell-surface proteins with cytoskeletal components and proteins involved in cytoskeletal dynamics. Mutations in this gene are associated with neurofibromatosis type II which is characterized by nervous system and skin tumors and ocular abnormalities.

NRAS: 1 Assay
The most important NRAS mutation in brain cancer occurs at codon 61.

PIK3CA: 3 Assays
The most frequently occurring PIK3CA mutations mainly belong to two classes: gain-of-function kinase domain activating mutations and helical domain mutations that mimic activation by growth factors.

PTEN: 12 Assays
The most commonly detected PTEN loss-of-function mutations are due to either truncation (p.R233* and p.R130*) or point mutations causing phosphatase inactivation (p.R130 and p.R173 mutations).

View a table of the mutations, associated COSMIC IDs and assay numbers, by clicking �Mutation Table� above on the right.

Related Biologies
Brain Cancer
Gene Resource List
Format
pdf
FileSize
137KB
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Name qBiomarker™ Somatic Mutation PCR Array Human Brain Cancers (SMH-025AA)

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