Counting PCR: An Alternative To Digital PCR For Absolute Quantification | 17086
Journal of Biotechnology & Biomaterials
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Anew method called ?Counting PCR? has been invented that analyzes the shape of a PCR curve to reveal the absolute
copy number of DNA at cycle zero. The method works with any qPCR instrument and has applications for absolute
quantification of viral load, gene expression, and copy number variation. Previously, qPCR has required each plate to be
calibrated using a dilution series of standards of known concentration. Alternatively, researchers have resorted to using qPCR
for relative quantification using cycle thresholds, C
, which are difficult to interpret especially when comparing results from
different laboratories. These shortcomings of traditional qPCR have prompted the development of digital PCR technologies,
which do not require standards for absolute quantification, but which tend to be expensive and low-throughput relative to
qPCR. The method of Counting PCR has been incorporated into a new program called ?
?, which automatically
computes the absolute copy number for every qPCR reaction on the plate without requiring a standard curve. This talk will
provide insights into the mechanism of PCR and reveal the quantized nature of PCR.
John Santa Lucia Jr. is Professor of Chemistry at Wayne State University and he is the co-founder, President and CEO of DNA Software, Inc. He is a world expert in
the thermodynamics and kinetics of DNA hybridization and folding. He has also done work in the fields of RNA 3D structure prediction and in the structural biology
of bacterial ribosomes using NMR spectroscopy. He has published more than 50 papers and his work has been cited more than 5000 times. His thermodynamic
parameters are used in numerous software packages for PCR design.
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