MONDAY, OCTOBER 7
PRE-CONFERENCE SHORT COURSE
9:00 am - 12:00 pm
Digital PCR Experiment Design and Primer
Jim Huggett, B.Sc. (Hons), Ph.D., Science Leader, Nucleic Acid Metrology, Molecular & Cell Biology, LGC
Growing Droplet Digital PCR Applications: Multiplexing, Linkage Analysis and EvaGreen
Dianna Maar, Ph.D., Senior Scientist, Applications, Bio-Rad Laboratories
Over the last two years the QX100 has enabled users to measure DNA and RNA targets without a standard curve and with a new level of precision and sensitivity. Currently, ddPCR is being recognized for its strength in detecting and measuring copy number variable genes, rare single-nucleotide polymorphisms, rare species and low-fold gene expression. Here I will discuss some of these current applications as well as the expanding applications for ddPCR™. Growing applications include micro RNAs, single-cell gene expression, gene linkage, multiplexing and EvaGreen applications.
GOING DIGITAL: WHAT CAN IT DO?
1:25 pm Chairperson's Opening Remarks
Philipp Angenendt, Ph.D., CTO, Inostics GmbH
» KEYNOTE PRESENTATION
1:30 Digital PCR: A Paradigm Shift in Molecular Measurement
Digital PCR (dPCR) offers a unique ability to perform sensitive molecular measurement without the need for a standard curve. By converting the real-time PCR analogue signal to a binary output, measurement is simplified considerably. This also results in improved precision, facilitating the measurement of smaller differences, and the format leads to more sensitive detection of minority mutations. This presentation will discuss the benefits as well as disadvantages of dPCR, discuss the digital MIQE guidelines and offer a prediction of how dPCR may impact molecular measurement in the future.
2:00 What Validation is Required for Digital PCR Measurements?
Leonardo Pinheiro, Ph.D., Senior Scientist, Bioanalysis Group, National Measurement Institute, Australia
One of the major strengths of digital PCR is the direct counting approach which can provide precise, quantitative measurements without reference to a calibrant. However, the accuracy of data depends on successful amplification from each template molecule and, like real-time PCR, assay optimization is still critical. The requirements for validation of digital PCR assays will be discussed in relation to applications such as copy number variation, rare mutant detection and target quantification.
2:30 Technical Explanation and Theory of Digital PCR: What Can Go Wrong and Why?
Ross Haynes, Biological Science Technician, Biochemical Science Division, National Institute of Standards and Technology
This presentation will briefly go over how digital PCR works,take an intuitive look at Poisson statistics, and focus on possible measurement issues. These measurement issues illustrate that some prior knowledge of the material may be required to make good measurements. Understanding the details of how digital PCR works will help users intelligently design experiments, troubleshoot, and analyze results.
3:00 Refreshment Break with Exhibit and Poster Viewing
3:45 Digital PCR: A Historical Perspective
Alec Morley, Emeritus Professor, Haematology and Genetic Pathology, Flinders University
The first publication to use the term "digital PCR" appeared in 1999 but during the preceding decade there had been a series of publications which used the method either to study the properties of single molecules or to quantify single molecules. The rise and fall of these early studies will be reviewed
Digital PCR: Looking Back and Moving Forward with Alec Morley
4:15 Droplet Digital PCR Applications for Next-Generation Sequencing
Nicholas J. Heredia, Ph.D., Staff Scientist, Biological Sciences Group, Bio-Rad Laboratories Inc.
NGS platforms are currently being used to quantify rare species targets in samples. Heavy processing of the starting template in NGS workflows can result in biases of the representation of these targets. Here we will discuss Droplet Digital PCR applications for cross validating NGS data with respect to rare species targets, as well as the excellent power of ddPCR quantification of NGS libraries for optimal sequencing performance.
4:45 Welcome Reception in Exhibit Hall
5:15 Close of Day
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