A Quarter Century and the PCR Applied Techniques and the Fields of Use Are Still Increasing in Numbers -- Parameters for Successful PCR Primer Design -- MIQE-Compliant Validation of MicroRNA Biomarker Signatures Established by Small RNA Sequencing -- Enhanced Probe-Based RT-qPCR Quantification of MicroRNAs Using Poly(A)tailing and 5' Adaptor Ligation -- A Novel System to Discriminate HLA-C mir148a Binding Site by Allele-Specific Quantitative PCR -- Detection of Yellow Fever Virus by Quantitative Real-Time PCR (qPCR) -- Evaluation of the Abundance of Fungi in Wastewater Treatment Plants Using Quantitative PCR (qPCR) -- Early Detection of Fungal Plant Pathogens by Real-Time Quantitative PCR: The Case of Diplodia sapinea on Pine -- A General Protocol for Accurate Gene Expression Analysis in Plants -- Gene Expression Analysis in Bacteria by RT-qPCR -- Detection and Characterization of Circulating Tumor Cells by Quantitative Real-Time PCR -- Molecular Monitoring of Chronic Myeloid Leukemia -- Normalization in Human Gliomas Tissue -- qPCR Applications for the Determination of the Biological Age -- QuantStudio™ 12K Flex OpenArray® System as a Tool for High-Throughput Genotyping and Gene Expression Analysis -- Digital PCR and the QuantStudio™ 3D Digital PCR System.

This book expands upon the useful first edition by exploring classic Quantitative Polymerase Chain Reaction (qPCR) techniques as well as a number of recently developed applications. With the changes in instrumentation due to technological advances and the development of new reagents to fulfill ethical and legal issues, the qPCR field is now an up-to-date technology that indeed is widely used in research and clinical diagnostics. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Revised and authoritative, Quantitative Real-Time PCR: Methods and Protocols, Second Edition is an ideal guide to this expanding and vital field of study.