Quantitative Real-Time PCR: Methods and Protocols

ISBN-13: 9781493907328 / Angielski / Twarda / 2014 / 231 str.

Roberto Biassoni; Alessandro Raso

Opis

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.

Recenzje

From the book reviews:

"The book is a valuable collection of qPCR-related details that cover a wide range of PCR uses. Aimed mainly at researchers (wet lab biologists) with a solid PCR background, the book provides enough details for the novice too, who may find the numerous examples as well as the end notes in each chapter very useful." (Irina Ioana Mohorianu, zbMATH, Vol. 1295, 2014)

Spis treści

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