ddPCR™ Mutation Detection Assays

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ddPCR Mutation detection assays can be found and purchased on Bio-Rad.com/digital-assays. A wet-lab validated mutation detection assay is returned if available, or an assay is designed instantly for your target. Wet-lab validated mutation detection assays detect mutant DNA present at 0.1% in a background of WT DNA in one well. An even better sensitivity of 0.005–0.1% can generally be achieved by using multiple reaction wells. Browse list of wet-lab validated Mutation Detection Assays.

Mutation detection assays can be designed instantly for over 4 million mutations, listed in the COSMIC database, on Bio-Rad’s Digital Assay Site. Built by the experts in digital PCR at Bio-Rad's Digital Biology Center, Bio-Rad's Droplet Digital PCR Assays Portal takes into consideration everything from GC content and Tm, to primer placement and probe length, even genome specificity when designing an assay for you.

Key Benefits

• Wet-lab validated assays available for more than 200 targets
• NEW Dark Probe assays detect up to 4 mutations per well on the QX600™ and QX700™ ddPCR Systems*
• Wet-lab validated assays guaranteed to work when used as instructed
• Validation data available for review and download
• Assays designed instantly using variant identifier or sequence of any species
• Suitable for ctDNA, liquid biopsy, FFPE, and other tissue samples
• Available in both standard and Dark Probe configurations
• Compatible with the QX200**, QX ONE*, QX600, and QX700 ddPCR Systems
• Orderable in single-tube or 2-tube formats
• Positive control sequences provided

* For Research Use Only. Not for use in diagnostic procedures.
** For Research Use Only. In vitro diagnostic (IVD) option is available

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Dark Probe Assays: Detect More Mutations Per Well

Unlock Higher-Plex Mutation Panels on Your Existing ddPCR System

Dark Probe Mutation Detection Assays replace the WT fluorescent probe with a quencher-only (dark) probe, suppressing WT signal without consuming a fluorescent channel. This frees each channel for an additional mutation target, doubling your panel capacity in a single well.

Dark Probe Assay Benefits

• Compatible with the QX series ddPCR Systems, Dark Probe technology enables detection of up to 4 mutation targets plus up to 2 (QX600 System) or 3 (QX700 System) reference assays in a single well, compared with 2–3 targets using standard assays
• Duplex mutation detection on the QX200 ddPCR System: go from 1 to 2 targets per well; no hardware upgrade required
• Exceptional sensitivity: achieve 0.05–0.1% VAF at 40,000 copies of WT input per well
• Self-serve design: design your Dark Probe assays instantly via the Digital Assays Portal for any of 4 million+ mutation targets
• Reduce cost per data point: fewer wells, less sample, lower reagent spend per experiment
• Tested across mutation types: point mutations (G>A, A>G, G>T, G>C), insertions, deletions, and MNVs

Consistent Rare Mutation Detection Across the QX600 and QX700 ddPCR Systems

ddPCR Mutation Detection Assays deliver consistent %VAF measurements acrosss the QX600 and QX700 ddPCR Systems. Figure 1 shows EGFR deletion, EGFR L858R, and KRAS G12C detected at expected VAF levels from 10% down to 0.01%, enabling confident mutation panel results regardless of which ddPCR platform is in use.

ddPCR Mutation Detection · 6-Plex Sensitivity

Clear 1-D droplet separation and linear titration across platforms

Fig. 1. Multiplex mutation detection assays demonstrate consistent performance across QX Systems at 0.01–10% mutant g-block in 40,000 copies of WT gDNA. EGFR del (FAM/HEX), EGFR L858R (Cy5/Cy5.5), KRAS G12C (ROX/Atto590).

Packaging Options

ddPCR Mutation Detection Assays are available in single tube or two-tube formats.

Single-tube format

Single-tube ddPCR Mutation Detection assays are 20x concentrated, ready-to-use primer-probe mixes optimized for use with Bio-Rad ddPCR systems. Single-tube assays are orderable in sizes of 200, 1,000, or 2,500 x 20 µL reactions through the Digital Assays Portal.

• Standard single-tube: each assay contains 18 µM of each primer and 5 µM of each probe. Mutant and WT probes are pre-mixed for added convenience. The available channel pairings are: FAM (MUT) + HEX (WT), Cy5 (MUT) + Cy5.5 (WT), ROX (MUT) + ATTO 590 (WT)
• Dark Probe single-tube (NEW): contains a fluorescent mutant probe (FAM, HEX, Cy5, or Cy5.5) paired with a quencher-only dark WT probe for wild-type signal suppression. Dark Probe assays contain an elevated WT quencher-only probe concentration to ensure effective suppression even at high DNA input

Dark Probe: Single Tube Format

Two-tube formats

The mutant and WT assays are provided as separate 20x primer-probe mixes, with each assay containing either the mutant probe or the WT probe. Two-tube assays are orderable in sizes of 200, 1,000, or 2,500 x 20 µL reactions through the Digital Assays Portal.

Related Products

• ddPCR Supermix for Probes no dUTP
• ddPCR Supermix for Probes
• ddPCR Multiplex Supermix
• QX700 ddPCR Supermix for Probes
• ddPCR KRAS G12/G13 Screening Kit
• ddPCR KRAS Q61 Screening Kit
• ddPCR NRAS G12 Screening Kit
• ddPCR NRAS G12/G13 Screening Kit
• ddPCR NRAS Q61 Screening Kit
• ddPCR BRAF V600 Screening Kit
• ddPCR EGFR Exon 19 Deletions Screening Kit

Resources

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  Droplet Digital PCR Applications Guide

  Read in-depth information about setting up experiments for ddPCR applications.

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  Rare Mutation Detection Best Practices Guidelines

  Learn best practices for performing mutation detection experiments.