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Custom qBiomarker Somatic Mutation PCR Arrays

For rapid and accurate profiling of somatic mutations using a custom gene panel

  • Profiling of somatic mutations using a custom gene panel
  • Simple real-time PCR procedure
  • Probe-based assays with high sensitivity and wide dynamic range
  • Optimized for robust performance on most PCR instruments
  • Master mix included

Custom qBiomarker Somatic Mutation PCR Arrays are translational research tools that allow rapid and accurate profiling of somatic mutations using a custom gene panel.

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Custom qBiomarker Somatic Mutation PCR Arrays

PCR plate with a custom panel of mutation assays

 337022 Varies

Custom 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.


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Profiling of common cancer cell lines for somatic mutation status.

200 ng gDNA from wild-type or cancer cell lines had 85 mutations (x axis) profiled on the Human EGFR qBiomarker Somatic Mutation PCR Array. Each detected mutation is indicated as a peak. All previously identified mutations in the EGFR pathway in these cell lines were confirmed.

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Sensitivity of qBiomarker Somatic Mutation PCR Arrays with FFPE samples.

The Human EGFR Pathway qBiomarker Somatic Mutation PCR Array profiled 85 mutations (x axis) in 4 FFPE cell line samples, a wild-type FFPE placenta sample control, and 9 lung adenocarcinoma FFPE tissue samples of primarily (8 out of the 9 samples) Caucasian origin with mixed gender and smoking status (samples along y axis). The array detected BRAF and KRAS mutations in the FFPE MDAMB- 231 cell line (shown as peaks), which were independently confirmed using the QIAGEN KRAS PCR Kit and Pyrosequencing. One KRAS somatic mutation and one EGFR somatic mutation were also detected in 2 FFPE lung adenocarcinoma samples.

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Principle of ARMS technology.
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High degree of sensitivity.

A dilution series of mixed genomic DNA samples derived from A375 cells (BRAF V600E positive) and wild type genomic DNA were tested for BRAF V600E with or without whole genome amplification (WGA). The sensitivity of this assay was determined to be 1–2%.

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 are ideal tools for the detection of mutations in cell lines or fresh or FFPE research samples. Sensitive and accurate evaluation of mutation status 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 master mix, 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 tool for 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

Custom qBiomarker Somatic Mutation PCR Arrays are highly suited for the rapid and accurate profiling of mutations for a custom panel of genes.

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