MAPK, KRAS, BRAF Genomic DNA (gDNA)

OncoREF

Applied StemCell Inc’s MAPK genomic DNA (gDNA) reference standards represent biologically-relevant controls that can be directly incorporated into your sample processing workflows in order to optimize your protocols, evaluate assay sensitivity and specificity, and analyze the impact of workflow changes on downstream analysis. They represent ideal materials for both assay development and routine monitoring of assay performance.

The MAPK Genomic DNA Reference Standards are extracted from ASC’s panel of isogenic MAPK mutation cell lines with 50 recurrent pathway-activating mutations in the EGFR, KRAS and BRAF genes, based on data from the Sanger Institute’s COSMIC database.

Key Features of the MAPK Series gDNA Reference Standards:

  • Most comprehensive MAPK mutation panel on the market
  • Well-characterized colorectal cancer cells lines: EGFR (RKO), KRAS (RKO), BRAF (HCT116)
  • Paired, isogenic wild-type cell lines to serve as an ideal control
  • Footprint-free, homozygous mutations
  • Reference cell lines are expanded from single-cells, ensuring maximum homogeneity
  • Available in multiple formats, including slides, scrolls, and full FFPE blocks
Need FFPE standards using your own cell lines?  Contact us for more details.

About the MAPK Pathway

The MAPK signaling pathway encompasses a series of signal transduction events that flow from the engagement of EGFR at the outer cell membrane, through KRAS, BRAF, MEK, and ERK.  MAPK signaling ultimately results in transcriptional activation of key genes that promote cellular proliferation, survival, differentiation, motility, and angiogenesis[1].  As such, the MAPK pathway is one of the most frequently activated pathways in cancer[2], and several drugs have pharmacogenomics profiles that depend upon the MAPK mutational status[3].


MAPK-pathway

To learn more about our FFPE Reference Standards and other diagnostic products, WATCH our WEBINAR!

Technical Details

Consistent Source of Biorelevant Specimens

Genomic DNA reference standards perform efficiently as reference material for qPCR applications:

  • Ability to detect mutations using qPCR platforms
  • Amplification efficiency close to 1
  • Idea for qPCR assay development and validation
TECHNICAL-MAPK-genomicDNA-2-qPCR-AJH

Support Materials

ONCOREF™ Reference Standards: Application of CRISPR/Cas9 to the Generation of Isogenic Cell Lines and Reference Materials (October 2016)

2016-Oncoref-webinar

 

 

 

 

 

 

 

 

 

CRISPR/Cas9 is rapidly enabling the development of new tools for enhancing our understanding oncogenic mutations in cancer. In order to aid in advancing cancer diagnosis and treatment, Applied StemCell has recently engineered a series of 40 isogenic cell lines that feature diverse mutations in the MAPK pathway. These mutant lines are available as isogenic pairs for applications in lead compound discovery, or as FFPE and nucleic acid reference materials for assay development. This webinar will focus on ASC’s efforts in developing these research tools, as well as applications of the materials for the advancement of cancer research.

Highlights of this talk:

  • Overview of molecular reference materials 
  • Workflow and QC for ONCOREF™ cell line generation
  • Advantages of CRISPR-engineered molecular reference standards
  • Applications of reference materials in assay development
  • Q & A
FAQ

1. How are your cell lines generated?

Answer: ASC uses CRISPR/Cas9 technology to enable precision genome editing in its MAPK mutation panel series.  This technology allows for footprint-free gene modification, meaning that you don’t have to worry about the presence of selection markers or other genomic footprints during the development of your assay technologies.

2. How do you validate mutational status in your cell lines? 

Answer: All of our MAPK mutation panel cell lines are expanded from single clones.  This ensures maximum homogeneity of the genetic profile.  After clonal expansion, we confirm the mutational status of the cell line using Sanger sequencing.

3. Do you offer mixtures of mutant and wild-type DNA?

Answer: ASC does not currently provide off-the-shelf cell line mixtures, but we can generate custom FFPE blocks to meet your particular research needs through our custom FFPE cell line services.

References
  1. Akinleye A, Furqan M, Mukhi N, Ravella P, Liu D (2013) MEK and the inhibitors: from bench to bedside. J. Hematol. Oncol. 6, 1–11
  2. Santarpia L, Lippman SM, El-Naggar AK (2012) Targeting the MAPK–RAS–RAF signaling pathway in cancer therapy, Expert Opin. Ther. Targets 16, 103–119
  3. Relling MV, Evans WE (2015) Pharmacogenomics in the clinic, Nature 526, 343–350
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