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Hervé Pouzoullic Life Technologies |
Abstract
Digital PCR is an extremely promising method for increasing accuracy, precision and sensitivity of nucleic acid quantification beyond the capabilities of traditional real-time qPCR. This is done by combining PCR assays with single molecule sensitivity with a system to partition a sample into a set of reactions which number close to, or more than, the total number of target molecules in that sample. The combination of these functions makes it possible to calculate the number of target molecules present by counting the number of reactions with or without amplification. Many novel applications are enabled by this new approach, including reference free absolute quantification and high sensitivity rare-allele quantification. We will be presenting a live demonstration of the QuantStudio™ 3D Digital PCR System, a new digital PCR platform from Life Technologies™. This chip based system enables collection of up to 20,000 data points per sample run. The workflow has been optimized for simplicity, minimizing hands-on time, minimizing the risk of sample cross contamination, and minimizing sample loss. In this workshop, we will review the basics of digital PCR theory and practice, discuss the capabilities of the QuantStudio™ 3D Digital PCR System, and demonstrate the steps involved in a full digital PCR experiment, including chip loading, running, imaging and data analysis.
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