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Ovarian Cancer Mutation PCR Array

Human
 
qBiomarker Somatic Mutation PCR Array: Human Ovarian Cancer
The Human Ovarian Cancer qBiomarker Somatic Mutation PCR Array is a translational research tool that allows rapid, accurate, and comprehensive profiling of the somatic mutations in human ovarian cancer samples in the following key genes: BRAF, CTNNB1/beta-catenin, ERBB2, FOXL2, GNAS, KIT, KRAS, NRAS, PIK3CA, PTEN, and P53. These mutations warrant extensive investigation to enhance the understanding of carcinogenesis and identify potential drug targets. The utility of somatic mutation status information in identifying key signaling transduction disruptions has been demonstrated in numerous research studies. For example, the mutation status of the EGFR and KRAS genes can predict the physiological response to certain drugs targeting these molecules. The Human Ovarian Cancer qBiomarker Somatic Mutation PCR Array, with its comprehensive content coverage, is designed for studying mutations in the context of ovarian cancer and has the potential for discovery and verification of drug target biomarkers for this cancer type and other cancer types in which these mutations have been identified. This array includes 83 DNA sequence mutation assays designed to detect the most frequent, functionally verified, and biologically significant mutations in human ovarian cancer. 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 2600 ovarian cancer samples. The simplicity of the product format and operating procedure allows routine somatic mutation profiling in any research laboratory with access to a real-time PCR instrument.

 

Assay Functional Annotations How It Works References Resources
 
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BRAF: 1 Assay
The most important BRAF mutation in ovarian cancer leads to increased kinase activity, the p. V600E mutation.

CTNNB1: 9 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.

ERBB2: 2 Assays
The most frequently identified ERBB2 activating mutations cluster in the ERBB2 kinase domain region.

FOXL2: 1 Assay
This mutation lies in wing 2 of the forkhead domain, a divergent component of the domain's secondary structure with unknown function, but the mutation does seem to inhibit the protein's pro-apoptotic function.

GNAS: 1 Assay
Mutations in this gene result in pseudohypoparathyroidism type 1a (PHP1a), which has an atypical autosomal dominant inheritance pattern requiring maternal transmission for full penetrance.

KIT: 3 Assays
The most frequently identified KIT gain-of-function mutations include the D816V point mutation, the exon 11 (juxtamembrane domain) deletion and point mutations, an exon 9 insertion mutation, and exon 13 point mutations.

KRAS: 10 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.

NRAS: 1 Assay
The most important NRAS mutation in ovarian cancer occurs at codon 12.

PIK3CA: 7 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: 3 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).

TP53: 45 Assays
The most frequently detected somatic mutations in TP53 are largely composed of DNA-binding domain mutations which disrupt either DNA binding or protein structure.

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

 

Assay Functional Annotations How It Works References Resources
 

Overview of the qBiomarker Somatic Mutation PCR Array / Assay Protocol

Overview of the qBiomarker Somatic Mutation PCR Array / Assay Protocol.
The procedure involves DNA extraction (QIAGEN QIAamp DNA Mini Kit or FFPE Tissue Kit is recommended), an optional amplification (QIAGEN REPLI-g kit or REPLI-g UltraFast kit is recommended) step for DNA isolated from fresh samples, qPCR detection on qBiomarker Somatic Mutation PCR Arrays or Assays, and data analysis (using the qBiomarker Somatic Mutation Data Analysis Template). An optional DNA sample QC step immediately before the detection array or assay setup allows the user to qualify the DNA samples.

Principle of Mutant Discrimination with ARMS®

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Assay Functional Annotations How It Works References Resources
 
  1. The molecular genetic basis of ovarian cancer and its roadmap towards a better treatment. Despierre E, Lambrechts D, Neven P, Amant F, Lambrechts S, Vergote I. Gynecol Oncol. 2010 May;117(2):358-65.
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Assay Functional Annotations How It Works References Resources
 

User Manual qBiomarker Somatic Mutation PCR Array System (PDF)
Data Analysis qBiomarker Somatic Mutation PCR Array Data Analysis Software
Application Data Detection Limits, Cancer Pathways
FAQ Frequently Asked Questions about Somatic Mutation Assays and Arrays
Webinar qBiomarker Somatic Mutation Analysis: Real-World Application Data
Slide Presentation> Presentation about qBiomarker Somatic Mutation Assays and Arrays (PDF)
Scientific Poster A Novel Tool for Pathway-Focused Cancer Mutation Profiling (PDF)
Presented at American Association for Cancer Research 2011
White Paper Rapid and accurate cancer somatic mutation profiling with the qBiomarker Somatic Mutation PCR Array (PDF)
Product Profile For screening biology-focused panels of gene mutations (PDF)

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