Overview
The ddPLEX EGFR/KRAS/BRAF Mutation Detection Kit* provides highly sensitive, reproducible, quantitative results for detecting key non-small cell lung cancer (NSCLC) markers in plasma or tumor tissue samples.
Targeting 37 actionable variants covering key EGFR, KRAS, and BRAF markers, this comprehensive kit includes a mutation assay, total quantification assay, positive control, and reagents for 120 reactions. Assay sets and positive controls are also available separately to support flexible laboratory workflows.
When paired with Bio-Rad’s QX600™ Droplet Digital™ PCR (ddPCR™) System* and using the recommended controls and guidelines, labs can process up to 42 unknown samples per plate and complete sample loading to data generation in under 8 hours, accelerating oncology research with confidence and precision.
* For Research Use Only. Not for use in diagnostic procedures.
Features and Benefits
- High sensitivity: detect key NSCLC mutations at 0.025%-0.1% VAF in a single well
- Accelerated turnaround: achieve same-day results with a streamlined workflow
- Versatile sample input: compatible with plasma or formalin-fixed paraffin embedded (FFPE) samples for greater flexibility
- Built-in quality assurance: includes internal and positive controls; positive control also available separately.
- Enhanced detection power: patented wild-type suppression chemistry boosts sensitivity
- Reliable performance: powered by ddPCR technology using the QX600 ddPCR System and QX Manager Software (SE v2.1 or higher) for exceptional reproducibility and specificity
Note: Variant Allele Frequency
Gene Coverage
| EGFR | KRAS | BRAF | |||||||
|---|---|---|---|---|---|---|---|---|---|
| Exon 18 | Exon 19 |
|
Exon 21 | Exon 2 | Exon 15 | ||||
| G719A G719S G719C |
Deletions | Insertions | S768I | T790M | C797S in cis/trans | L858R | L861Q | G12C | V600E |
Performance
Example data with separated clusters across 6 channels (FAM, HEX, Cy5, Cy5.5, ROX, and ATTO590) on QX600 ddPCR System.
Figure 1. ddPLEX EGFR/KRAS/BRAF Mutant Assay
Figure 2. ddPLEX EGFR/KRAS/BRAF Total Quantification Assay
This assay can also identify whether C797S variants are present in cis configuration (on the same allele) or trans configuration (on a different allele) relative to T790M.
Figure 3. T790M/C797S configuration discrimination
This figure displays clusters for the T790M, T790M+C797S T>A (cis and trans), T790M+C797S G>C (cis and trans), and C797S templates in the ROX/ATTO590 channels.
- Plot A: T790M
- Plots B and C: C797S variants
- Plot D: cis-configured T790M and C797S T>A
- Plot E: cis-configured T790M and C797S G>C
- Plots F and G: trans-configured T790M and C797S
- Plot H: T790M, cis-configured T790M and C797S T>A, cis-configured T790M and C797S G>C, and C797S G>C
Analytical Sensitivity
Table 1. Summary of LoB and LoD results for the ddPLEX EGFR/KRAS/BRAF Mutant Assay (≤ 0.1% LoD)
| Target | Limit of Blank (cp/µL) | Limit of Detection (Variant Allele Frequency) |
Limit of Detection (Mutant Copies in 40,000 Total) |
|---|---|---|---|
| EGFR ex18, G719X | 0.51 | 0.025–0.05% | 10 |
| EGFR ex19, del | 0.05 | 0.05% | 20 |
| EGFR ex20, ins | 0.05 | 0.025–0.05% | 10-20 |
| EGFR ex20, S768I | 0.08 | 0.025% | 10 |
| EGFR ex20, C797S | 0 | 0.025% | 10 |
| EGFR ex20, T790M | 0.56 | 0.10% | 40 |
| EGFR ex20, L858R | 0.07 | 0.025% | 10 |
| EGFR ex20, L861Q | 0 | 0.025% | 10 |
| KRAS G12C | 0.05 | 0.025% | 10 |
| BRAF V600E | 0 | 0.025% | 10 |
* Samples with concentration below the LoB and/or 2 or fewer positive droplets were considered as negative.
Analytical Specificity
Samples with each of the EGFR, KRAS, and BRAF variants at 0.1% in a background of wild-type cell line DNA were tested at an input amount of 132 ng in the ddPLEX EGFR/KRAS/BRAF mutant assay. Values below the LOB and/or 2 or fewer positive droplets were considered as negative.
Figure 4. Analytical specificity of the ddPLEX EGFR/KRAS/BRAF Mutant Assay
Assay Linearity
The ddPLEX EGFR/KRAS/BRAF Mutation Detection Kit produces linear results from 20%–0.1% VAF (800 copies to 4 copies per reaction) in a background of 4,000 copies of human genomic DNA with R2>0.98.
Figure 5. Assay linearity of the ddPLEX EGFR/KRAS/BRAF Mutation Assay
Applications & Uses
This Research Use Only kit is used for the detection, discrimination, and quantification of key EGFR, KRAS, and BRAF mutations for NSCLC research, from cfDNA in plasma and/or DNA in FFPE tissue samples.
Included Components
The ddPLEX EGFR/KRAS/BRAF Mutation Detection Kit (17010491) includes the following components:
| Material Number | Description |
|---|---|
| 12023484 |
ddPLEX EGFR/KRAS/BRAF Mutation Detection Assay
|
| 12023522 | ddPLEX EGFR/KRAS/BRAF Positive Control (1 x 100 µL) |
| 12005909 | ddPCR Multiplex Supermix (2 x 600 µL) |
| 12012171 | Dithiothreitol (DTT) (2 x 1,000 µL) |
The four components above can also be purchased individually.
Configuration Options (or Instrument Compatibility)
The ddPLEX EGFR/KRAS/BRAF Mutation Detection Kit should be used with the QX600 ddPCR System and either the Manual Droplet Generator or Automated Droplet Generator. Equipment, key consumables, and reagents necessary for the QX600 ddPCR System are listed below (additional items may be needed, please see the User Guide for more details).
Key Droplet Generation & Reader Equipment, Consumables and Reagents
| Description | Catalog Number |
|---|---|
| QX600 Droplet Reader | 12013328 |
| Automated Droplet Generator | 1864101 |
| QX200 Manual Droplet Generator | 1864002 |
| PTC Tempo Thermal Cycler | 12015392 |
| PCR Plate Heat Seal | 1814040 |
| PX1 PCR Plate Sealer | 1814000 |
| ddPCR Plates 96-well Semi-skirted | 12001925 |
| ddPCR Droplet Reader Oil | 1863004 |
| ddPCR Buffer Control for Probes (2X) | 1863052 |
Reagents & Consumables for Automated Droplet Generation Only
| Description | Catalog Number |
|---|---|
| Automated Droplet Generator Oil for Probes | 1864110 |
| DG32 Automated Droplet Generator Cartridges | 1864108 |
| Pipet Tips for Automated Droplet Generator | 1864120 |
| Pipet Tip Bins for Automated Droplet Generator | 1864125 |
Reagents & Consumables for Manual Droplet Generation Only
| Description | Catalog Number |
|---|---|
| Droplet Generator Oil for Probes | 1863005 |
| Droplet Generator DG8 Cartridge Holder | 1863051 |
| DG8 Cartridges for QX100/QX200 Droplet Generator | 1864008 |
| DG8 Gaskets for QX100/QX200 Droplet Generator | 1863009 |
Related Products
- BRAF V600 ddPCR Multiplex Mutation Screening Kit
- KRAS G12/G13 ddPCR Multiplex Mutation Screening Kit
- EGFR Exon19 Deletions ddPCR Multiplex Mutation Kit
- ddPLEX ESR1 Mutation Detection Kit
- ddPCR™ Expert Design Assay: EML4-ALK, Human, Homo sapiens
- ddPCR™ Expert Design Assay: ROS1 mRNA Fusions, Human, Homo sapiens
- ddPCR™ Expert Design Assay: RET mRNA Fusions, Human, Homo sapiens
Related Categories
| SKU Number | Product Name |
|---|---|
| 17010491 | ddPLEX EGFR/KRAS/BRAF Mutation Detection Kit |
| 12023484 | ddPLEX EGFR/KRAS/BRAF Mutation Detection Assays
|
| 12023522 | ddPLEX EGFR/KRAS/BRAF Positive Control (1 x 100 µL) |
| 12005909 | ddPCR Multiplex Supermix (2 x 600 µL) |
| 12012171 | Dithiothreitol (DTT) (2 x 1000 µL) |
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The ddPLEX EGFR/KRAS/BRAF Mutation Detection Kit detects and quantifies 37 key non-small cell lung cancer (NSCLC) variants and their wild-type counterparts, which allows for calculation of variant allele frequencies in both plasma and tumor tissue samples.
ddPLEX EGFR/KRAS/BRAF Mutation Detection Assays detect and quantify 37 key non-small cell lung cancer (NSCLC) variants and their wild-type counterparts, which allows for calculation of variant allele frequencies in both plasma and tumor tissue samples.
The ddPLEX EGFR/KRAS/BRAF Positive Control is used with the ddPLEX EGFR/KRAS/BRAF Mutation Detection Assays to detect key non-small cell lung cancer (NSCLC) variants and allows for calculation of variant allele frequencies in both plasma and tumor tissue samples.
Accessories
Kits & Reagents
Pkg of 30, cartridges for use with the Automated Droplet Generator in the QX600/QX200 AutoDG Droplet Digital PCR System
70 ml (10 x 7 ml), oil for use with droplet generator in the QX600/QX200 Droplet Digital™ PCR Systems
Pkg of 24, cartridges for use with the QX200 Droplet Generator
Pkg of 24, gaskets for use with QX200 Droplet Generator
Downloads
-
QX Manager Software Standard Edition, Version 2.1QX-MANAGER-V.2.1-STD
QX Manager Software Standard Edition, Version 2.1
-
ddPLEX EGFRKRASBRAF MT TQ Assays Template CSVSOFT-ddPLEX-MT-TQ-TEMP
A simple, structured CSV for entering key digital assay details to ensure consistent and efficient setup across platforms.