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Tuesday, February 11
7:00 am Registration and Morning Coffee
9:15 Refreshment Break in the Exhibit Hall with Poster Viewing
10:25 Chairperson’s Remarks
10:30 Counting Copies: Digital Distributions and Math for Melting
Robert Palais, Ph.D., Associate Professor, Math Dept, Utah Valley University; Research Professor, Pathology Dept, University of Utah
Joint work with CT Wittwer, L Zhou, S Sundberg, Z Dwight
We will discuss two mathematical algorithms that can be used to quantify DNA copy number variation with increased accuracy. One permits rapid exact evaluation of cumulative probability distributions having one or more degrees of freedom, for sampling with or without replacement. Another normalizes high-resolution melting peak of the target with respect to that of a reference after ensuring the initial copy ratio is preserved during PCR.
11:00 Eprobe: New Fluorescence Probe for the Combination of Real-Time PCR and Melting Curve Analysis
Takeshi Hanami, Ph.D., LSA Technology Development Unit, Omics Science Center, RIKEN
Eprobe is a hybridization-sensitive fluorescent probe that only shows strong fluorescence signals upon hybridization to a complementary DNA strand. This talk will describe how the probe combines real-time PCR monitoring and melting curve analysis as well as how we achieved multiplex detection by using multicolor Eprobes. The combination provides powerful means for new mutation detection assays in a single-tube reaction.
11:30 PACS: PCR-Activated Cell Sorting
Adam R. Abate, Ph.D., Assistant Professor, Bioengineering and Therapeutic Sciences, California Institute for Quantitative Biosciences (QB3), University of California, San Francisco
We have developed a microfluidic system that allows individual cells to be analyzed and sorted based on the outcomes of single-cell PCR reactions. Our system encapsulates each cell in a microdroplet, lyses the cell, and performs RT-PCR to detect gene transcripts, SNPs, or small non-coding RNAs of interest. Unlike FACS, PACS requires no antibodies and can differentiate among cells based on transcriptional variation. The technology has broad applications in basic research and medical diagnostics, including in cancer, immune function, and microbiology.
12:00 pm Whole-Body Scanning PCR; A Highly Sensitive Method to Study the Biodistribution of Therapeutic Oligonucleotides
Iwan Beuvink, Ph.D., Protein Production and Antibodies, NIBR Biologics Center, Novartis Institutes for Biomedical Research
Efficient tissue-specific delivery is one of the crucial factors required for the successful development of therapeutic oligonucleotides. Whole-body scanning PCR is a platform that can be used to study the tissue-homing properties of novel delivery strategies by visualizing the in vivo biodistribution of the formulated therapeutic oligonucleotide. The platform relies on the local tissue-extraction from a mouse whole-body section followed by the conversion of target-specific qPCR signals into an image.
12:30 Session Break
12:40 Luncheon Presentations (Sponsorship Opportunities Available) or Lunch on Your Own
1:40 Refreshment Break in the Exhibit Hall with Poster Viewing
2:15 Chairperson’s Remarks
Fred Russell Kramer, Ph.D., Professor, Department of Microbiology and Molecular Genetics, Public Health Research Institute, New Jersey Medical School
2:20 Application of Digital PCR for the Analysis of RNA
Rebecca Sanders, Researcher, Nucleic Acid Metrology, Molecular Biology, Science and Technology Division, LGC
2:50 Digital PCR Developments We'd Like to See
N. Reginald Beer, Ph.D., Medical Diagnostics Initiative Leader, Center for Micro and Nanotechnologies, Lawrence Livermore National Laboratory
The development of digital PCR has led to many exciting applications that benefit from the technique's ability to isolate, amplify, and detect rare or numerically-disadvantaged targets. Not surprisingly, this created early interest in medical diagnostics where the ability to detect rare mutants among a wild type background is critical. More widespread adoption, however, would benefit from additional advances that increase the technology's value proposition. In this talk we will discuss several trends that could broaden dPCR's appeal.
3:20 Picoliter Droplet-Based Digital PCR for Molecular Diagnostics
Valerie Taly, Ph.D., Group Leader/CNRS Researcher, Université Paris-Descartes
Picoliter droplet-based digital PCR allows the highly sensitive and quantitative detection of rare cancer markers within complex mixtures of DNA like patient samples. We will present the development of droplet multiplex procedures for the quantitative detection of the seven most frequent KRAS mutant alleles as well as wild type sequences and clinical applications of these procedures for patient treatment management. The results of two clinical trials involving metastatic cancer patients will be presented.
3:50 Liquid Biopsy – Blood-Based Molecular Testing in Oncology
Frank Diehl, Ph.D., Vice President, Lab Operations, Research & Development, Sysmex Inostics
The use of blood for molecular testing in oncology opens up new, non-invasive possibilities for the management of cancer patients in the context of therapy selection, response prediction, real time follow-up, and resistance monitoring.
4:05 Sponsored Presentation (Opportunity Available)
4:20 Valentine’s Day Celebration in the Exhibit Hall with Poster Viewing
5:20 Breakout Discussions in the Exhibit Hall
These interactive discussion groups are open to all attendees, speakers, sponsors, & exhibitors. Participants choose a specific breakout discussion group to join. Each group has a moderator to ensure focused discussions around key issues within the topic. This format allows participants to meet potential collaborators, share examples from their work, vet ideas with peers, and be part of a group problem-solving endeavor. The discussions provide an informal exchange of ideas and are not meant to be a corporate or specific product discussion.
Very Fast (Extreme) PCR
Moderator: Carl Wittwer, M.D., Ph.D., Professor, Pathology, University of Utah
- What are the uses of PCR in < 1 min?
- Is extreme PCR practical?
- How can you maintain specificity with high primer and polymerase concentrations?
PCR Quality Control Considerations
Moderator: Rebecca Sanders, Researcher, Nucleic Acid Metrology, Molecular Biology, Science and Technology Division, LGC
- Special considerations for dPCR
- Publishing accurate, clear data
- Ensuring reliable reagents
- Strategies for experimental design
Additional Breakout Discussions to be Announced
6:30 Close of Day
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