Main description:

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.

Contents:

1. Twenty Years of qPCR: A Mature Technology?

Alessandro Raso and Roberto Biassoni

2. Minimum Information Necessary for Quantitative Real-Time PCR Experiments

Gemma Johnson, Afif Abdel Nour, Tania Nolan, Jim Huggett, and Stephen Bustin

3. Selection of Reliable Reference Genes for RT-qPCR Analysis

Jan Hellemans and Jo Vandesompele

4. Introduction to Digital PCR

Francisco Bizouarn

5. mRNA and micro RNA Purity and Integrity: The Key to Success in Expression Profiling

Benedikt Kirchner, Vijay Paul, Irmgard Riedmaier, and Michael W. Pfaffl

6. Mediator Probe PCR: Detection of Real-Time PCR by Label Free Probes and a Universal Fluorogenic Reporter

Simon Wadle, Stefanie Rubenwolf, Michael Lehnert, Bernd Faltin, Manfred Weidmann, Frank Hufert, Roland Zengerle, and Felix von Stetten

7. Absolute Quantification of Viral DNA: The Quest for Perfection

Domenico Russo and Mauro Severo Malnati

8. 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

Francesco Broccolo

9. Real-Time PCR Detection of Mycoplasma pneumoniae in the Diagnosis of Community-Acquired Pneumonia

Eddi Di Marco

10. A Sensible Technique to Detect Mollicutes Impurities in Human Cells Cultured in GMP Condition

Elisabetta Ugolotti and Irene Vanni

11. Real-Time Quantification Assay to Monitor BCR-ABL1 Transcripts in Chronic Myeloid Leukemia

Pierre Foskett, Gareth Gerrard, and Letizia Foroni

12. A Reliable Assay for Rapidly Defining Transplacental Metastasis Using Quantitative PCR

Samantha Mascelli

13. Circulating Cell-Free DNA in Cancer

Pamela Pinzani, Francesca Salvianti, Claudio Orlando, and Mario Pazzagli

14. Gene Expression Analysis by qPCR in Clinical Kidney Transplantation

Michael Eikmans, Jacqueline D.H. Anholts, and Frans H.J. Claas

15. Post-Transcriptional Regulatory Networks: From Expression Profiling to Integrative Analysis of mRNA and Micro RNA Data

Swanhild U. Meyer, Katharina Stoecker, Steffen Sass, Fabian J. Theis, and Michael W Pfaff

16. Clinical Applications Using Digital PCR

Francisco Bizouarn

17. Developing Non-Invasive Diagnosis for Single Gene Disorders: The Role of Digital PCR

Angela N. Barrett and Lyn S. Chitty