BRAF V600E Reference Standard, 5%

Synonyms: B-Raf, BRAF1

Forma: FFPE

The BRAF V600E Reference Standard is a highly-characterized, biologically-relevant quality control material used to assess the performance of assays that detect somatic mutations, such as Sanger and qPCR sequencing assays.

With this product you are able to:

• Evaluate of workflow integrity from pre-analytical DNA extraction to post-analytical bioinformatics
• Analyze the sensitivity of your workflow
• Optimize and validate new cancer panels and routinely monitor the performance of your assay

Horizon FFPE standards eliminate the variability associated with patient-derived reference standards, and avoid the hassle of sourcing, characterizing, and documenting your own cell line mixes. The standard is provided at an allelic frequency of 5%, and may be diluted using the corresponding wild type standard to generate a lower allelic frequencies for limit of detection studies and validation of your pre-analytical phase of the workflow.

Horizon’s Base-Seq products are all derived from human cell lines. Using a proprietary method of mild fixation and homogenous paraffin embedding, we generate highly-reproducible and consistent FFPE curls. With any commercially available FFPE extraction protocol, our standards yield high molecular weight DNA. This format ensures that they may applied to a wide-range of assays including qPCR, Sanger sequencing, next-generation sequencing, mass array, and more.

Technical Data

DNA Base Change: GTG→GAG

NCBI Reference Assembly SNP: rs113488022

Cosmic ID: COSM476

Fixation Method: 4% Formalin

Section Size: 15µm or 20µm

Cell Density: 3 x 10⁸ cells per block. Approx. 3.5 x 10⁵ cells per section

Expected DNA Yield: ≥ 400 ng using Promega Maxwell LEV Plus Extraction kit

Product Information

Intended Use: For routine performance monitoring (Research Use Only)

Unit Size: 1 FFPE Section per vial

Extractable DNA: ≥ 400 ng per section using Promega Maxwell LEV Plus FFPE DNA Extraction kit

General Information

Allelic Frequency: 5%

Storage: 4˚C

Expiry: 48 months from the date of manufacture

Cell Line Background: HCT116

Quality Control

Allelic Frequency: Droplet Digital PCR™

Genotype: Sanger sequencing of locus specific PCR

Quality: Agarose gel electrophoresis

Quantification: Quantifluor™

Genotype: BRAF (V600E/+)

Characterization
BRAF V600E Reference Standard - Chromatogram
Chromatograph showing heterozygosity for the B-Raf V600E mutation within B-Raf exon 15 (SNP accession number: rs113488022)

Primers
    Primer    Sequence                                             Size
    Forward    CTGGGCCTACATTGCTAAAAT              513
    Reverse    AGTTGAGACCTTCAATGACTTTCTA
    Sequencing    AGTTGAGACCTTCAATGACTTTCTA    --