Digital PCR (dPCR) and Quantitative PCR (qPCR) Solutions

Quantitative PCR (qPCR or real-time PCR) and RT-qPCR (reverse transcription quantitative PCR) methods measure the abundance of specified nucleic acid sequences in a sample. qPCR combines PCR amplification with a fluorescent probe that reacts to the presence of a specific DNA or RNA sequence. This simple approach is a powerful investigational tool with broad applications in healthcare, research, quality assurance, and many other fields.

Our expert scientists can assist at virtually any stage of your qPCR or RT-qPCR project. We accept fresh frozen tissue, blood, buccal swabs, cell pellets, purified RNA, cDNA, or gDNA as starting material. Please see our sample submission guidelines for details on sample types, minimum starting materials, and recommended QC parameters.

We can accept CLIA or GLP-grade qPCR projects on a case-by-case basis. Please contact our CLIA team via email at CLIA@azenta.com if you're interested in clinical-grade services.

Digital PCR (dPCR) is a new method of nucleic acid quantification that partitions a sample into thousands of nanoliter-sized droplets or wells, each of which undergoes PCR amplification. The resulting data can be used to determine the absolute quantity of the target nucleic acid sequence in the sample. 

A: qPCR amplifies a target sequence in the presence of a fluorescent dye until it reaches detectable levels. dPCR partitions these reactions into thousands of droplets or wells and each partition is amplified and measured independently. This enables more precise quantification of the target nucleic acid in a sample, down to single-copy resolution. 

ddPCR involves partitioning a sample into thousands of water-in-oil droplets using a specialized droplet generator. Chip-based dPCR, on the other hand, involves partitioning a sample into thousands of wells on a microfluidic chip. Both techniques have similar principles, but the methods of partitioning differ. One advantage of digital PCR methods is that the number of available partitions does not vary between reactions in the same experiment. 

Digital PCR is a targeted approach that quantifies the absolute amount of a known DNA sequence in a sample, whereas NGS is an unbiased approach that sequences millions of DNA fragments simultaneously to identify and quantify a sample. 

dPCR offers increased precision, reproducibility, and sensitivity over qPCR, RT-qPCR, and other available nucleic acid quantitation methods. Digital PCR is also more robust in the presence of PCR inhibitors, allowing even challenging samples to be quantified effectively. dPCR can also be used to detect rare targets in complex samples, such as low-abundance transcripts in total RNA. 

dPCR has broad applications in research and development, clinical diagnostics, biotechnology, and beyond. dPCR excels in copy number variation (CNV), detection of low-level mutations or rare transcripts, and is used in the evaluation of viral titer as well as quantification of viral vector copy number (VCN) of treated cells for AAV, lentivirus, and retrovirus in clinical research. 

Experimental design and expertise in dPCR quantitation are the most pivotal factors for success in any dPCR experiment. Other key considerations include the quality and quantity of starting material, reaction conditions, and primer/probe design. 

Absolute quantification determines the exact number of copies of a target nucleic acid in a sample, whereas relative quantification measures the fold change in target nucleic acid expression between two samples. 

Our expert scientists can assist at virtually any stage of your dPCR project.  We accept fresh frozen tissue, blood, buccal swabs, cell pellets, purified RNA, cDNA, or gDNA as starting material. Please see our sample submission guidelines for details on sample types, minimum starting materials, and recommended QC parameters. 

Our team can design and optimize dPCR assays.  Please contact molgen@azenta.com to find out more about our custom design services. 

By working with Azenta, researchers and companies can benefit from customized and optimized dPCR solutions that can improve accuracy, sensitivity, and efficiency, while reducing variability and development time. 

Our team is composed of expert scientists who can discuss your project goals and determine how Azenta can best assist you with design and customization services for dPCR.  Please contact us via email at molgen@azenta.com to set up a technical consultation session.