12. Nanoliter High-Throughput PCR for DNA and RNA Profiling
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The increasing emphasis in life science research on utilization of genetic and genomic information underlies the need for
high-throughput technologies capable of analyzing the expression of multiple genes or the presence of informative single nucleotide
polymorphisms (SNPs) in large-scale, population-based applications. Human disease research, disease diagnosis, personalized
therapeutics, environmental monitoring, blood testing, and identification of genetic traits impacting agricultural practices,
both in terms of food quality and production efficiency, are a few areas where such systems are in demand. This has stimulated
the need for PCR technologies that preserves the intrinsic analytical benefits of PCR yet enables higher throughputs without
increasing the time to answer, labor and reagent expenses and workflow complexity. An example of such a system based on a
high-density array of nanoliter PCR assays is described here. Functionally equivalent to a microtiter plate, the nanoplate
system makes possible up to 3,072 simultaneous end-point or real-time PCR measurements in a device, the size of a standard
microscope slide. Methods for SNP genotyping with end-point TaqMan PCR assays and quantitative measurement of gene expression
with SYBR Green I real-time PCR are outlined and illustrative data showing system performance is provided.
Series: Methods in Molecular Biology | Volume: 496 | Pub. Date: Oct-24-2008 | Page Range: 161-174 | DOI: 10.1007/978-1-59745-553-4_12
Subject: Genetics/Genomics
Key Words: 5′-exonuclease assay - TaqMan PCR - SNP genotyping - nanofluidic - high-throughput genotyping - SYBR Green I real-time PCR - nanoliter PCR - quantitative PCR
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