Molecular Assays

Molecular Biological Assays for Accurate Analysis and Diagnosis

Molecular assays are commonly used in support of drug development, particularly with newer drug modalities such as cell and gene therapies. BioAgilytix leverages real-time polymerase chain reaction, quantitative PCR or qPCR, and Droplet Digital PCR (ddPCR) as part of our extensive molecular technology suite to deliver bioanalytical solutions to support discovery, preclinical, and clinical programs.

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BioAgilityx’s PCR expertise

Our scientific team offers a breadth and depth of expertise in the development, optimization, and validation of complex molecular assays using qPCR and ddPCR, collaborating with your team to ensure your project is a success. Our laboratories are equipped with the latest generation of PCR equipment, including Applied Biosystems QuantStudio 7 Flex or Pro with both 96- and 384-well fast-cycle options, as well as TaqMan Array Cards and BioRad’s droplet digital PCR systems, QX200 and QXOne. 

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PCR for molecular assay support at any phase of drug development

Common PCR-based methods supported by BioAgilytix include:

  • Custom, real-time qPCR
  • Gene expression analysis using TaqMan assays
  • SNP genotyping analysis and copy number variation
  • Mutation detection 
  • PK of RNA-based therapeutics
  • Gene and cell therapy support (biodistribution/persistence, viral shedding)

Selecting qPCR or ddPCR for high-quality molecular results

Molecular assays in support of bioanalytical testing can be performed on either qPCR or ddPCR. Real-time PCR (RT-qPCR or qPCR) is often considered the gold standard in molecular analysis for detecting nucleic acids. However, ddPCR is a newer technology that can have distinct advantages over traditional PCR methods in specific situations.  

Applications for qPCR

qPCR monitors DNA amplification in real-time by continually measuring the signal of fluorescent molecules (chromophores) activated during the PCR process. Modern qPCR instrumentation allows for the simultaneous measurement of multiple, unique DNA amplifications in a single reaction vessel, making it possible to quantitate multiple targets per sample. This translates into lower sample volumes and reduced reagent usage, creating substantial overall time and cost savings. BioAgilytix uses the latest Quant Studio platforms that are scalable with both 96- or 384-well blocks. The qPCR platform can obtain 10 logs of linear range and is useful with assays that require wide dynamic ranges.

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Applications for ddPCR

ddPCR is inherently more precise than traditional PCR as ddPCR amplifies individual DNA fragments rather than from bulk sampled DNA. In ddPCR, samples are fractionated into thousands of individual droplets. Individual DNA fragments in the sample are thereby partitioned or distributed within the droplets which allows direct counting of the number of DNA molecules within a sample (i.e. the number of droplets in the sample that contain DNA).  This method provides high precision at low copy numbers. BioAgilytix uses ddPCR QX200 and QX One for absolute quantification of target DNA or RNA (cDNA) molecules without the need for a standard curve. QX ONE ddPCR System is used for its multiplexing capabilities to simultaneously detect multiple targets. This system seamlessly integrates a standard ddPCR workflow of droplet generation, thermal cycling, droplet reading, and analysis into a robust platform. The walk-away automation of the QX ONE allows our scientists to provide robust results.