Since becoming commercially available in 2011, digital PCR (dPCR) has been gaining in popularity as it enables nucleic acid quantification with superior accuracy and precision compared to quantitative PCR (qPCR). In brief, the technology consists of the following steps:
- The PCR reaction mixture is partitioned into thousands of water-in-oil droplets with target and background DNA randomly distributed among the reactions
- The target DNA is amplified by PCR using standard thermal cycling with fluorescent dyes or probes
- Each reaction provides a fluorescent positive or negative signal indicating that target DNA was present or absent in partitioning
- The fraction of positive droplets is used to calculate the target DNA concentration using Poisson correction
With data acquisition taking place at the end of the reaction, and since absolute concentrations can be determined, data can be easily interpreted and compared across different samples and experiments, especially in a clinical setting. Additionally, dPCR doesn’t suffer from the inherent complications of qPCR that result from the requirement of a standard curve and excels in the quantification of low abundant genes or small differences.
This webinar will describe the intrinsic potential of droplet digital (ddPCR) in clinical research for three different applications:
- Rare variant analysis in plasma samples
- Small changes in alternative splicing
- CAR-T studies
During the talk, the featured speaker will analyze the rationale for dPCR selection and present relevant case studies.
Jan Hellemans, PhD, Co-founder and Chief Technology Officer, Biogazelle
Jan Hellemans is founder and CTO of Biogazelle, a Belgian service provider exploiting RNA to improve healthcare. He is an expert in qPCR and dPCR technologies and co-author of the (digital)MIQE guidelines. He led the development of Bio-Rad’s PrimePCR assays and supports Biogazelle’s service team in designing and validating qPCR/ddPCR assays for clinical research.Message Presenter
Who Should Attend?
This webinar is aimed towards those working in a pharma, biotech, CRO or public organization, with relevant job titles including:
- Principal Investigators
- (Senior) Scientists
- Study Directors
- Heads of Research
Relevant areas of interest include:
- Quantification of nucleic acids, e.g. gene copy numbers
- Gene and micro-RNA expression analysis
- Oncology, e.g. rare allele detection or adoptive cell therapies
- Quantification of pathogen loads
What You Will Learn
In this webinar, participants will learn:
- A comparison of dPCR and qPCR, including advantages and disadvantages of each technique
- Why partitioning increases the variant fraction in the sample
- Best practices for limit of detection (LOD) and limit of quantification (LOQ) determination
- Elements of a validation plan for a clinical trial study
Biogazelle is a CRO specializing in high-value applications to support pharmaceutical research, clinical trials, and diagnostic test development. To accelerate the development of small molecules, RNA targeted drugs and adoptive cell therapies, we apply a suite of genomic and transcriptomic technologies, to find and validate RNA biomarkers and to assess efficacy, safety, and toxicity. We hold a unique forefront position in the application of quantitative PCR, digital PCR, and dedicated RNA sequencing workflows on precious clinical samples such as liquid biopsies and FFPE tissues. Our laboratories are ISO/IEC 17025:2005 accredited and PCR based services can be performed in GCLP compliance.