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Digital PCR Conference: Applications and Advances - Day 2

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8:00-8:25 am Registration and Morning Coffee

8:25 Chairperson’s Opening Remarks

Kerry R. Emslie, Ph.D., Bioanalysis Group Manager, National Measurement Institute, Australia


Kenneth Kinzler 8:30 Digital Genomics and the Study of Cancer - Download Podcast 

Kenneth W. Kinzler, Ph.D., Professor, Oncology; Director, Ludwig Center at Johns Hopkins University, The Johns Hopkins Kimmel Cancer Center

Digital enumeration of genetic material is finding growing utility in both basic research and clinical applications. Digital PCR, digital expression and digital karyotyping approaches will be discussed in the context of the study of human cancer.

9:15 PANEL: Implementing Digital PCR in the Lab

Moderator: Kerry R. Emslie, Ph.D., Bioanalysis Group Manager, National Measurement Institute, Australia

Panelists: Andy Watson, Chief Marketing Officer and Head of Global Sales, RainDance Technologies

George Karlin-Neumann, Ph.D., Sr. Director of ddPCR Assay Development, Bio-Rad Digital Biology Center

10:00 Coffee Break with Exhibit and Poster Viewing


10:30 Chairperson’s Remarks

Daniel Weisenberger, Ph.D., Assistant Professor, Research, USC Epigenome Center

• • • Technology • • • 

RainDance Technologies10:35 Single Molecule Digital PCR for Sensitivity Detection of Cancer Biomarkers

Darren R. Link, Ph.D., Co-Founder and Vice President, Research and Development, RainDance Technologies, Inc.  - Download Podcast 

Somatic mutations within tumoral DNA are highly specific biomarkers that distinguish cancer cells from normal cells. While these biomarkers are likely to be useful for the diagnosis, prognosis and treatment of patients, they require a highly sensitive technique that can discriminate tumor-specific mutations in a large excess of non-mutated DNA. The RainDance Digital PCR solution utilizes up to 10 million reactions per lane to detect one mutant amongst 250,000 wild-type molecules.

• • • Application • • • 

11:05 Droplet PCR for Personalized Cancer Healthcare

Pierre Laurent-Puig, M.D., Ph.D., Director, Molecular Basis of the Response to Xenobiotics, Descartes University

Tumor cells are characterized by the presence of specific gene alterations, which can be useful for the monitoring of cancer patients through the development of biomarkers such as a free circulating tumor DNA in different body fluids. Such an approach led us to the development of highly sensitive methods because the tumor DNA is buried in an excess of normal DNA. The digital droplet PCR enabled us to reach this goal through an easy and reproducible method, which had been validated in a series of metastatic colorectal cancer patients for KRAS mutations.

• • • Technology • • • 

Bio-Rad11:35 High Precision, Ultra-Sensitive Droplet Digital PCR (ddPCR)

George Karlin-Neumann, Ph.D., Director of Scientific Affairs, The Digital Biology Center, Bio-Rad Laboratories

ddPCR extends the range of reliable detection and quantitation of biomarkers, especially those at low levels. Sample partitioning into droplets followed by PCR and flow cytometry detection allows the direct counting of target molecules as fluorescent droplets. The high precision and sensitivity of the system will be illustrated by examples drawn from rare event detection, CNV, absolute quantitation, gene expression and emerging ones.

• • •  Application • • • 

12:05 pm Digital Detection of Rare Mutations and Tumor Infiltrating T Cells: Clinical Application in Cancer and Disease

Jason H. Bielas, Ph.D., Assistant Member, Molecular Diagnostics Program, Fred Hutchinson Cancer Research Center (FHCRC); Affiliate Assistant Professorship, Department of Pathology, University of Washington

12:35 Luncheon Presentations (Opportunities Available, contact Jay Mulhern;


1:35 Chairperson’s Remarks

Christina Fan, Ph.D., Senior Scientist, ImmuMetrix LLC

1:40 Approaches to Single Cell Analysis

Wendy Winckler, Ph.D., Scientific Director, Genomics Platform, The Broad Institute

We have developed methods in collaboration with Fluidigm to genotype mutations and measure gene expression from individual cells. We apply these techniques to the study of cell lineage and clonal evolution in CLL.

2:10 DNA Methylation Analysis Using Digital MethyLight Technology

Daniel Weisenberger, Ph.D., Assistant Professor, Research, USC Epigenome Center

DNA methylation changes are frequently observed in all human cancers, and can be also identified in patient blood. Here we describe the development of Digital MethyLight technology, a digital PCR application of MethyLight, sensitively detecting DNA methylation in primary tissues and fluids. We have identified and quantitated single-molecule, cancer-specific DNA hypermethylation events in plasma and serum samples from patients with cancer. This technology has promise in cancer detection and surveillance.

2:40 Digital Amplification of DNA Molecules and Metaphase Chromosomes with Applications in Prenatal Diagnosis

Christina Fan, Ph.D., Senior Scientist, ImmuMetrix LLC

This presentation will cover our work in microfluidic digital PCR for aneuploidy detection, and the extension of digital PCR of DNA molecules to microfluidic digital multiple strand displacement amplification (MDA) of metaphase chromosomes from single cells. Digital MDA of chromosomes enables whole-genome haplotyping of human individuals, and the application in noninvasive prenatal diagnosis would be also be described.

3:10 Refreshment Break with Exhibit and Poster Viewing


3:40 Chairperson’s Remarks

Christina Fan, Ph.D., Senior Scientist, ImmuMetrix LLC

3:45 Large-Scale Droplet Arrays for Digital Microfluidics and Digital PCR

Abraham P. Lee, Ph.D., William J. Link Professor and Chair, Department of Biomedical Engineering; Director, Micro/Nano Fluidics Fundamentals Focus (MF3) Center, University of California, Irvine

To increase the dynamic range of digital PCR, a larger number of droplet (emulsion) reactors are required to detect a wider range of DNA concentrations. In this presentation, a large-scale (up to 1-million) droplet array is described that is enabled by droplet microfluidics that also enables monodispersed reactor volumes and stacking in 3-D for higher droplet density per unit area. In addition, our platform enables nucleic acid processing with real-time fluorescence measurements, which provide temporal information about PCR amplification.

4:15 Real-Time Droplet DNA Amplification with a New Tablet Platform

Anubhav Tripathi, Ph.D., Co-Director, Center for Biomedical Engineering, Brown University

This talk presents a novel droplet-based tablet platform for temporal polymerase chain reaction (PCR) in microliter droplets. The simple design of the device does not require extensive processing or external equipment, which allows for greater ease of use and integration as a point-of-care diagnostic.

4:45 Sponsored Presentations (Opportunities Available, contact Jay Mulhern;

5:15 Digital Nucleic Acid Amplification in a Sample Self-Digitization (SD) Chip

Daniel T. Chiu, Ph.D., A. Bruce Montgomery Professor of Chemistry, Endowed Professor in Analytical Chemistry, University of Washington

This presentation will describe the development and operation of our sample self-digitization (SD) chip and its application in digital nucleic acid amplification. I will describe the realization of digital LAMP in the SD chip as well as the implementation of digital PCR using the SD chip.

5:45 Close of Conference

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