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Signature® KRAS/BRAF Mutations*

The Signature® KRAS/BRAF Mutations assay is a research tool for the rapid detection of mutations in genomic DNA extracted from human cell lines or fresh, frozen or fixed tissues. KRAS and BRAF are oncogenes involved in the epidermal growth factor receptor (EGFR) signaling pathway that controls cell proliferation, differentiation and apoptosis. Mutations in the KRAS and BRAF oncogenes are frequently found in human cancers, such as colorectal cancers and non-small cell lung cancers.

The qualitative assay simultaneously detects:

  • 6 KRAS mutations in codons 12
  • 6 KRAS mutations in codon 13
  • 1 BRAF mutation in codon 600

KRAS/BRAF RUO Workflow
The Signature® reagents are adapted for use on the Luminex® 100 IS™ or 200™ Systems.

Assay Features:

Streamlined workflow, protocol and reagents optimized for the molecular lab

Compatible with a broad range of genomic DNA input and type*

  • FFPE, biopsies, FNA, macro- or micro-dissected cells
  • 6 KRAS mutations in codon 13
  • Recommended input for optimal performance: 5-20 ng genomic DNA

Qualitative assay with simple data interpretation:

  • A specimen is positive for a given mutation if signal ≥ validated cut off
  • Recommended research use cutoff: 400-500 MFI

Analytical sensitivity* of about 1% demonstrated with genomic DNA and model FFPE cell line dilutions

Includes internal endogenous control (EC) to assess sample DNA quality/purity

External positive and negative plate controls provided independently to assess the validity of the amplification, hybridization and detection steps in each run

Assay Controls

KRAS Mutation Controls BRAF Mutation Controls

The control set provides negative and positive controls for both panels:

  • G12V Positive Control
  • G13V Positive Control
  • BRAF V600E Positive Control
  • KRAS/BRAF Pooled Control (all mutations detected)

Assay Sensitivity

KRAS Mutation Analysis Sensitivity BRAF Mutation Analysis Sensitivity

Analytical sensitivity* of at least 1% with KRAS mutant positive cell line DNA diluted in KRAS WT cell line DNA

Assay Specificity

KRAS/BRAF Mutation Specificity

Excellent analytical specificity* with no cross detection between KRAS and BRAF

  • The HCT1116 cell line is KRAS G13D positive and wild type BRAF
  • The HT29 cell line is KRAS wild type and BRAF V600E positive

 


*Research Use Only. Not intended for use in diagnostic procedures.

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