MONDAY, OCTOBER 15
Pre-Conference Short Course*
9:00 am-12:00 pm
• An Introduction to Digital PCR
Jim Huggett, Ph.D., Science Leader, Nucleic Acid Metrology, LGC
• Principles of Digital PCR and Measurement Issues: The Certification of Cytomegalovirus Standard Reference Material (SRM 2366) as a Model for Future SRMs
Ross Haynes, Biological Science Technician, Biochemical Science Division,
National Institute of Standards and Technology
• Billion Reactions. Digital Answers: The Next Generation of PCR is Here
Michael Samuels, Principal Research Scientist & Scientific Liaison, Raindance Technologies, Inc.
* Separate Registration Required
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12:00-1:00 pm Main Conference Registration
12:55 Chairperson’s Opening Remarks
N. Reginald Beer, Ph.D., Medical Diagnostics Initiative Leader, Center for Micro and Nanotechnologies, Lawrence Livermore National Laboratory
1:00 Digital PCR: Moving Out of the Lab and onto the Bench
N. Reginald Beer, Ph.D., Medical Diagnostics Initiative Leader, Center for Micro and Nanotechnologies, Lawrence Livermore National Laboratory
Digital PCR provides extremely accurate quantitation as compared to traditional standard curve methods but at what cost? Originally, dPCR instruments were slow, time-consuming, proprietary, and expensive- all of which hindered the adoption of the technique to all but academic applications. Recently, the marketplace has begun to catch up- with only familiarity and the cost of operation hindering further adoption. In this talk we will discuss the benefits of dPCR, its most appropriate applications, and future trends driving the field.
1:30 Validation of Digital PCR: Generating Reliable Data
Kerry R. Emslie, Ph.D., Bioanalysis Group Manager, National Measurement Institute, Australia
Digital PCR provides a level of precision in target DNA quantification which is difficult to achieve using other PCR platforms — but how can the user be confident the data is accurate and reliable? Critical factors such as sample preparation, sample homogeneity, reaction volume measurements, thermal cycler uniformity, template GC content and methylation can all effect data quality. This presentation will discuss the role of reference materials and the use of orthogonal methods to verify the accuracy of dPCR.
2:00 qPCR CopyCountTM: Mechanistic qPCR Quantification as an Alternative to Digital PCRJohn SantaLucia, Jr., Ph.D., President & CEO, DNA Software, Inc.qPCR CopyCountTM is a software tool that fits raw qPCR data using mechanism-based modeling to determine highly accurate relative and absolute DNA concentrations without the use of standards, without specialized equipment, and without digital PCR. qPCR CopyCountTM has applications in non-invasive diagnostics for copy number variation, gene expression analysis, and genotyping in cancer screening, fetal testing, and agriculture.
2:15 Microfluidic Tools for PCR and Digital PCR
Bruce K. Gale, Ph.D., Associate Professor and Director, State of Utah Center of Excellence for Biomedical Microfluidics, Department of Mechanical Engineering, University of Utah
Microfluidics has been used to develop a rapid and specific PCR and digital PCR tools. This presentation will specifically address PCR methods such as gradient-based, continuous flow PCR and disk based digital PCR. These tools have been used to complete PCR and digital PCR with high resolution melting analysis in less than 8 minutes. Results of combining these systems with automated microfluidic extraction systems will also be presented.
2:45 Microreactor Arrays for Digital PCR, Sequencing, and Single Cell Analysis
Peter A. Sims, Ph.D., Assistant Professor, Department of Biochemistry and Molecular Biophysics, Columbia Initiative in Systems Biology, Columbia University Medical Center
Microfluidic devices fabricated using soft lithography have enabled a variety of biological applications over the last decade, including digital PCR. I will describe a simple and versatile microfluidic device, the reversibly sealable microreactor array, which serves as the basis for both massively parallel sequencing and digital PCR platforms. I also will discuss recent efforts to develop a single cell analysis tool based on this system to directly link conventional phenotypic observations of individual cells with their transcriptomes.
3:15 Digital PCR for Rapid Non-Invasive and Prenatal Aneuploidy Detection Through Genetic Analysis of Circulating Trophoblastic Cells Isolated by ISETPatrizia Paterlini M.D., Ph.D., Professor, Cell Biology & Oncology and Director, INSERM; Founder, Rarecells SAS & Rarecells USA, Inc.Circulating Fetal Trophoblastic Cells (CFTC) are consistently and reliably isolated from maternal blood using ISET technology starting from the 5th week of gestation. Since CFTC provide fetal DNA not mixed with maternal DNA, they represent the optimal target for digital PCR allowing the prenatal and non-invasive detection of Trisomy 21 and other aneuploidies through a simplified and rapid workflow.
3:45 Refreshment Break with Exhibit and Poster Viewing
4:15 The Advantages and Future of Digital PCR
Jim Huggett, Ph.D., Science Leader, Nucleic Acid Metrology, LGC
Developments in microfluidics and emulsion chemistries have made digital PCR an easy to perform technique that builds on the capabilities of real time PCR while also offering a simple approach. With continuous advances to increase throughput and reduce cost, dPCR has the potential to have a major impact on molecular analysis for research and applied applications like diagnostics. This talk will focus on the current potential of dPCR and where the future is likely to take it, especially in the context of the recent major technological advance of next-generation sequencing.
4:40 The Advantages and Future of Next-Generation Sequencing
Shawn C. Baker, Ph.D., CSO, BlueSEQ
The field of next-generation sequencing has been rapidly expanding and improving over the past several years and looks to continue doing so for the foreseeable future. Concomitant with this improvement has been the rapid expansion of applications addressable by next-gen sequencing. The talk will focus on how well the various applications fit with the different platforms and how this will likely change with future advancements on the horizon. Special emphasis will be placed on those applications most likely to overlap with digital PCR, examples of which include minority target detection, copy number variation and absolute counting applications.
5:05 PANEL: Digital PCR vs. NGS
- Will sequencing technology mature to the point where digital PCR is unnecessary?
- Is there room for both technologies?
- How can the two work in tandem?
- Will cost of these technologies be prohibitive?
Panelists:
Jim Huggett, Ph.D., Science Leader, Nucleic Acid Metrology, LGC
Shawn C. Baker, Ph.D., CSO, BlueSEQ
5:30 Welcome Reception with Exhibit and Poster Viewing
6:30 Close of Day One