PrimePCR™ ddPCR™ Mutation Assay: BRAF WT for p.V600R, Human

PrimePCR Primer Assays for Real-Time PCR oligo primer pair tube for SYBR Green gene expression

ddPCR probe assay designed for mutation detection. Probe assays consist of unlabeled PCR primers and a dual labeled fluorescent probe.

Fluorophore:   HEX
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Assay Information

Technology:   ddPCR
Assay Type   Probe
Application:   Mutation Detection
Unique Assay ID:   dHsaCP2000038
Chromosome Location:   7: 140453120-140453242question
Amplicon Length:   91
Cosmic ID   COSM474
Nucleotide Mutation Detection   WT for c.1798_1799GT>AG
Amino Acid Change   WT for p.V600R
Restriction Enzymes   MseI

Gene Information

This gene encodes a protein belonging to the raf/mil family of serine/threonine protein kinases. This protein plays a role in regulating the MAP kinase/ERKs signaling pathway which affects cell division differentiation and secretion. Mutations in this gene are associated with cardiofaciocutaneous syndrome a disease characterized by heart defects mental retardation and a distinctive facial appearance. Mutations in this gene have also been associated with various cancers including non-Hodgkin lymphoma colorectal cancer malignant melanoma thyroid carcinoma non-small cell lung carcinoma and adenocarcinoma of lung. A pseudogene which is located on chromosome X has been identified for this gene. [provided by RefSeq Jul 2008]

Gene Symbol:   BRAF
Gene Name:   v-raf murine sarcoma viral oncogene homolog B1
Aliases:   B-RAF1, BRAF1, FLJ95109, MGC126806, MGC138284, NS7, RAFB1
RefSeq:   NC_000007.13 NG_007873.1 NT_007914.15
Ensembl:   ENSG00000157764
Entrez:   673
UniGene:   Hs.550061
Chromosome Mapping   7q34

  Download Validation Data (.pdf)

Products used to generate validation data:

ddPCR Instrument QX100
ddPCR Supermix ddPCR Supermix for Probes (no dUTP)
Experimental Sample Horizon HCT116 (mut), Promega female human genomic DNA (WT)

%Fractional Abundance
Dilution series of mutant DNA in ~40,000 copies of wild type DNA background.

ddPCR Amplitude Scatter Plot (wild type + mutant)
Single-well data for mutant DNA spiked into wild-type DNA (~0.1%). Mutation assay (FAM, Channel 1) was duplexed with the corresponding wild-type reference assay (HEX, Channel 2).

ddPCR Amplitude Scatter Plot (wild type + mutant)

ddPCR Amplitude Scatter Plot (wild type)
Single-well data for wild-type DNA (no mutant). Mutation assay (FAM, Channel 1) was duplexed with the corresponding wild-type reference assay (HEX, Channel 2).

ddPCR Amplitude Scatter Plot (wild type)

Number Description Download
10033171 Instruction Manual, PrimePCR ddPCR Copy Number Variation (CNV) Assays, Rev A Click to download
10033487 PrimePCR ddPCR Mutation Detection Assays Product Insert, Rev B Click to download
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