EGFR Gene-Specific Multiplex Reference Standard (gDNA)

The Gene-Specific DNA Reference Standards are a highly-characterized, biologically-relevant quality control material used to assess the performance of NGS assays that detect somatic mutations. These standards are available specifically in either an EGFR Multiplex (12.5% allelic frequency range) or a KRAS Multiplex (16.7% allelic frequency range).

Next generation sequencing (NGS) platforms give researchers much greater power when profiling tumor samples. However, the use of NGS presents challenges surrounding sample variability, platform bias, and potential failure to detect biomarkers. Horizon’s standards are appropriate for any NGS library preparation including whole-genome, whole-exome, custom capture and targeted amplicon panels. Horizon has developed this Quality-Seq (Q-Seq) NGS Reference Standard range to support the development and continued validation of Next Generation Sequencing platforms. The Gene-Specific Multiplex Reference Standard portfolio covers multiple endogenous SNPs, insertions and deletions at defined allelic frequency ranges. The Gene-Specific Reference Standards cover EGFR or KRAS-specific mutations. These standards may be diluted with a matched wild type while the background mutations remain consistent. The Gene Specific Multiplex Standards are available in genomic DNA and FFPE format, which allows you to validate both the pre-analytical and analytical portion of your workflow. The FFPE format allows you to evaluate and optimize DNA extraction protocols without wasting precious patient samples. By multiplexing multiple gene-specific mutations in a single standard, the Gene-Specific Multiplex allows researchers to maximize the information obtained from their assay, and confirm that they are able to detect a range of potential mutants in a single gene.

With this product you are able to:

  • Analyze the sensitivity of your assay
  • Gain certainty of the limit of detection and limit of quantification
  • Optimize and validate new cancer panels and routinely monitor the performance of your assay

 

Technical Data

Genes Covered: EGFR

Allelic Frequencies: 12.5

Buffer: Tris-EDTA (10mM Tris-HCl, 1mM EDTA), pH 8.1

 

Product Information

Verified Mutations: The EGFR Multiplex Reference Standard has been manufactured using multiple engineered cell lines and mixed to generate multiple Allelic Frequencies in multiplex. An example of the allelic frequency by whole-exome sequencing for one batch of this product is reported in the table below.

Gene Variant (AA) CDS mutation GRCh38 co-ordinates Expected allelic frequency (%) NGS allelic frequency (%) NGS read depth
EGFR L861Q c.2582T>A 7:55191831 12.50% 11.41% 184
EGFR ΔE746-A750 c.2235_2249del 7:55174771 12.50% 8.99% 345
EGFR L858R c.2573T>G 7:55191822 12.50% 6.53% 199
EGFR T790M c.2369C>T 7:55181378 12.50% 11.83% 896

Presence confirmed in parental cell line

Gene Variant (AA) CDS mutation GRCh38 co-ordinates Expected allelic frequency (%) NGS allelic frequency (%) NGS read depth
BRAF V600E c.1799T>A 7:140753336 66.70% 65.45% 631
PIK3CA H1047R c.3140A>G 3:179234297 50.00% 49.65% 711
FGFR1 P150L c.542C>T 8:38428093 50.00% 52.56% 390
MLH1 L323M c.967C>A 3:37020392 50.00% 46.09% 1586
CDH1 N/A c.*110dup 16:68833601 50.00% b b

b – Variant type not called by variant caller (large insertion, CN variant, fusion, indel)

Unit Size: 1 µg

Concentration: 50 ng/µl

 

General Information

Storage: 4˚C

Expiry: See all product shelf life information

 

Quality Control

Allelic Frequency: Droplet Digital PCR™

Genotype: Sanger sequencing of locus specific PCR

Quality: Agarose gel electrophoresis

Quantification: Spectrophotometry (A260)