Quantitative Real-Time PCR: Methods and Protocols focuses on different applications of qPCR ranging from microbiological detections (both viral and bacterial) to pathological applications. Several chapters deal with quality issues which regard the quality of starting material, the knowledge of the minimal information required to both perform an assay and to set the experimental plan, while the others focus on translational medicine applications that are ordered following an approximate logical order of their medical application. The last part of the book gives you an idea of an emerging digital PCR technique that is a unique qPCR approach for measuring nucleic acid, particularly suited for low level detection and to develop non-invasive diagnosis. Written for the Methods in Molecular Biology series, most chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, laboratory protocols and tips on troubleshooting and avoiding known pitfalls.

Practical and authoritative, Quantitative Real-Time PCR: Methods and Protocols aims to aid researchers seeking to devise new qPCR-based approaches related to his or her area of investigation.

Includes cutting-edge methods and protocols in qPCR techniques Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt

Twenty Years of qPCR: A Mature Technology?.- Minimum Information Necessary for Quantitative Real-Time PCR Experiments.- Selection of Reliable Reference Genes for RT-qPCR Analysis.- Introduction to Digital PCR.- mRNA and micro RNA Purity and Integrity: The Key to Success in Expression Profiling.- Mediator Probe PCR: Detection of Real-Time PCR by Label Free Probes and a Universal Fluorogenic Reporter.- Absolute Quantification of Viral DNA: The Quest for Perfection.- A Multiplex Real-Time PCR-Platform Integrated into Automated Extraction Method for the Rapid Detection and Measurement of Oncogenic HPV Type-Specific Viral DNA Load from Cervical Samples.- Real-Time PCR Detection of Mycoplasma pneumoniae in the Diagnosis of Community-Acquired Pneumonia.- A Sensible Technique to Detect Mollicutes Impurities in Human Cells Cultured in GMP Condition.- Real-Time Quantification Assay to Monitor BCR-ABL1 Transcripts in Chronic Myeloid Leukemia.- A Reliable Assay for Rapidly Defining Transplacental Metastasis Using Quantitative PCR.- Circulating Cell-Free DNA in Cancer.- Gene Expression Analysis by qPCR in Clinical Kidney Transplantation.- Post-Transcriptional Regulatory Networks: From Expression Profiling to Integrative Analysis of mRNA and Micro RNA Data.- Clinical Applications Using Digital PCR.- Developing Non-Invasive Diagnosis for Single Gene Disorders: The Role of Digital PCR.