Nanoplate dPCR applications

Rare mutation detection (RMD) refers to detecting a sequence variant that is only present at a very low frequency in a pool of wild-type backgrounds (less than 1% or even 0.1%). Thus, for detecting and quantifying rare events, such as point mutations or single nucleotide polymorphisms (SNPs), a sensitive, accurate and precise method is necessary. The challenge is the discrimination between two highly similar sequences, of which one is significantly more abundant than the other.

Benefits of using nanoplate dPCR for detecting rare mutations: 

• Ability to load a large input reaction volume into 26,000 partitions, which substantially increases the chances of finding a rare target 

• Multiplexing for mutant and wild-type sequencing to detect low fractions of rare mutant molecules against an abundant wild-type background

Multiplex digital PCR for mitochondrial and genomic target copy number analysis

Explore a workflow for high-throughput analyses of target copy numbers in cultured cells. The process combines fast and accurate cell sorting from CellenONE to ensure an exact number of cells are used in downstream reactions. The samples are subjected to probe-based detection on the QIAcuity Digital PCR System with multiplexing of up to five targets in a single dPCR reaction with minimal optimization. The workflow achieves accurate absolute DNA quantification by dPCR for analysis of low abundance targets as well as multi-copy targets at single-cell level.

The increased precision of digital PCR can provide higher resolution in many aspects of gene expression analysis. It enables monitoring finer changes in expression levels, even less than two-fold. Digital PCR also provides a greater sensitivity when quantifying rare targets, or RNA from very limited starting material.

Emerging uses of digital PCR for gene expression analysis include:

• Studying methylation events
  
• Quantifying transcription levels
  
• Measuring changes in miRNA levels

Digital PCR enables a range of gene therapy applications, including adeno-associated virus (AAV) vector genome titer and lentiviral vector copy number measurements and CAR T-cell therapy development and manufacturing. It also helps validate the quality of your therapeutic product with confidence. This is critical when developing effective and reproducible cell and gene therapies while ensuring patient safety.

Discover how QIAcuity dPCR can generate the same level of accuracy and precision in viral titer quantification as the traditional ddPCR method at increased speed and overall higher throughput and scalability.