PCR Technology Current Innovations, Third Edition

PCR’s simplicity as a molecular technique is, in some ways, responsible for the huge amount of innovation that surrounds it, as researchers continually think of new ways to tweak, adapt, and re-formulate concepts and applications. PCR Technology: Current Innovations, Third Edition is a collection of novel methods, insights, and points of view that provides a critical and timely reference point for anyone wishing to use this technology. Topics in this forward-thinking volume include: The purification and handling of PCR templates The effect of the manufacture and purification of the oligonucleotide on PCR behavior Optimum buffer composition Probe options The design and optimization of qPCR assays Issues surrounding the development and refinement of instrumentation Effective controls to protect against uncertainties due to reaction variability Covering all aspects of PCR and real-time PCR, the book contains detailed protocols that make it suitable as both a reference and an instruction manual. Each chapter presents detailed guidelines as well as helpful hints and tips supplied by authors who are recognized experts in their fields. In addition to descriptions of current technology and best practices, the book also provides information about new developments in the PCR arena.

Template PreparationPre-PCR Processing Strategies; Johannes Hedman, Maria Lövenklev, Petra Wolffs, Charlotta Löfström, Rickard Knutsson, and Peter RådströmUse of a Liposomal Internal Control Vehicle for Whole-Process Quality Assurance of Nucleic Acid Amplification; Einar S. Berg and Tania NolanReaction ComponentsOligonucleotide Synthesis and Purification; Yannick Fillon, Theresa Creasey, Qin Xiang, and Carlos MartinezAugmenting PCR: Effects of Additives in the Polymerase Chain Reaction; Ernest J. MuellerReal-Time Fluorescent PCR by Labeled Primer with a Single Fluorescent Molecule; Hong Yan and Liu Xiao KunUsing Scorpion Primers for Genotyping; David WhitcombeLNA: Adding New Functionality to PCR; Søren M. Echwald, Ditte Andreasen, and Peter MouritzenModified dNTPs: A Toolbox for Use in PCR; Natasha Paul, Olga Adelfinskaya, Elena Hidalgo Ashrafi, Tony Le, and Sabrina ShoreInstrumentsThermocycler Calibration and Analytical Assay Validation; Mary Span, Marc Verblakt, and Tom HendrikxUltra-High-Speed PCR Instrument Development; Nick Burroughs and Emmanouil KarterisDropletization of Bioreactions: How Single-Cell Quantitative Analyses May Improve Gene Measurement and Cancer Diagnosis; Ehsan Karimiani, Amelia Markey, and Philip DayDesign, Optimization, QC, and StandardizationAssay Design for Real-Time qPCR; Gregory L. ShipleyPrimer Design for Large-Scale Multiplex PCR and Arrayed Primer Extension; Maido Remm, Kaarel Krjutškov, and Andres MetspaluDevelopment and Use of qPCR Assays for Detection and Study of Neglected Tropical and Emerging Infectious Diseases; Ashley R. Heath, Norha Deluge, Maria Galli de Amorim, and Emmanuel Dias-NetoMIQE: Guidelines for Reliable Design and Transparent Reporting of Real-Time PCR Assays; Tania Nolan and Stephen A. BustinData AnalysisHypothesis-Driven Approaches to Multivariate Analysis of qPCR Data; Anders Bergkvist, Azam Sheikh Muhammad, and Peter DamaschkeExperiment Design, Data Management, and Univariate Statistical Analysis of Gene-Expression Data Obtained by Real-Time Quantitative PCR; Ales Tichopad, Tzachi Bar, Malin Edling, Boel Svanberg, Tania Nolan, and Anders BergkvistBiomarker Discovery via RT-qPCR and Bioinformatical Validation; Christiane Becker, Irmgard Riedmaier, and Michael W. PfafflOptimization of Quantitative Real-Time PCR for Studies in Cartilage Mechanobiology; Oto Akanji, Donald Salter, and Tina ChowdhuryApplicationsEnriching DNA Sequences with Nucleotide Variation by Thymidine Glycosylases Combined with Suppression PCR; Xinghua Pan, Alexander E. Urban, and Sherman M. WeissmanChromosome Walking by Inverse PCR; Michael Gotesman, Selwyn A. Williams, Jorge A. Garcés, and Ray H. GavinExpression Profiling of MicroRNAs by Quantitative Real-Time PCR: The Good, the Bad, and the Ugly; Mirco Castoldi, Paul Collier, Tania Nolan, and Vladimir BenesDye-Based High-Throughput qPCR in Microfluidic Platform BioMark™; David Svec, Vendula Rusnakova, Vlasta Korenkova, and Mikael KubistaSurface Plasmon Resonance-Based Biosensor Technology for Real-Time Detection of PCR Products; Giordana Feriotto and Roberto GambariCurrent Innovations in the Development and Application of the qPCR in a Resource-Limited Setting; Clare Watt and Jim HuggettA Review of Isothermal Nucleic Acid Amplification Technologies; Gavin J. Nixon and Claire A. BushellProximity Ligation Assay for Protein Quantification; David W. Ruff, Mark E. Shannon, Shiaw-Min Chen, Simon Fredriksson, and Ulf LandegrenHigh-Resolution Melt Analysis; Einar S. Berg and Tania NolanIndex

Dr. Tania Nolan is an internationally recognized molecular biologist with a reputation for expertise in the field of mRNA quantification using RT-qPCR. She was a significant coauthor of the specialist textbook The A–Z of Quantitative PCR along with numerous other book chapters and peer-reviewed papers. She has presented several plenary lectures and chaired sessions at major international qPCR meetings. Stephen A. Bustin, BS(Mod) PhD FSB acquired his first qPCR instrument in 1997 and has published numerous peer-reviewed papers that describe and use this technology. He wrote and edited the A–Z of Quantitative PCR, universally acknowledged as the "qPCR Bible," edited The PCR Revolution, and has written a series of Definitive qPCR ebooks. He also led the international consortium that drew up the MIQE guidelines.