Quantitative Real-Time PCR

1. Front Matter

   Pages i-xiii

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2. A Quarter Century of PCR-Applied Techniques and Their Still-Increasing Fields of Use

   • Alessandro Raso, Roberto Biassoni

   Pages 1-4

3. Parameters for Successful PCR Primer Design

   • Stephen A. Bustin, Reinhold Mueller, Tania Nolan

   Pages 5-22

4. MIQE-Compliant Validation of MicroRNA Biomarker Signatures Established by Small RNA Sequencing

   • Veronika Mussack, Stefanie Hermann, Dominik Buschmann, Benedikt Kirchner, Michael W. Pfaffl

   Pages 23-38

5. Enhanced Probe-Based RT-qPCR Quantification of MicroRNAs Using Poly(A) Tailing and 5′ Adaptor Ligation

   • Valentin Vautrot, Isabelle Behm-Ansmant

   Pages 39-54

6. A Novel System to Discriminate HLA-C mir148a Binding Site by Allele-Specific Quantitative PC R

   • Priscilla Biswas, Eddi Di Marco, Mauro S. Malnati

   Pages 55-64

7. Detection of Yellow Fever Virus by Quantitative Real-Time PCR (qPCR)

   • Gisela Freitas Trindade, Sheila Maria Barbosa de Lima, Constança Britto, Alice Gomes Fernandes-Monteiro

   Pages 65-77

8. Evaluation of the Abundance of Fungi in Wastewater Treatment Plants Using Quantitative PCR (qPCR)

   • Paula Maza-Márquez, Elisabet Aranda, Jesús González-López, Belén Rodelas

   Pages 79-94

9. Early Detection of Fungal Plant Pathogens by Real-Time Quantitative PCR: The Case of Diplodia sapinea on Pine

   • Nicola Luchi, Alberto Santini, Francesca Salvianti, Pamela Pinzani

   Pages 95-104

10. A General Protocol for Accurate Gene Expression Analysis in Plants

    • Ellen De Keyser, Laurence Desmet, Magali Losschaert, Jan De Riek

    Pages 105-118

11. Gene Expression Analysis in Bacteria by RT-qPCR

    • Danilo J. P. G. Rocha, Thiago L. P. Castro, Eric R. G. R. Aguiar, Luis G. C. Pacheco

    Pages 119-137

12. Detection and Characterization of Circulating Tumor Cells by Quantitative Real-Time PCR

    • Francesca Salvianti, Filomena Costanza, Gemma Sonnati, Pamela Pinzani

    Pages 139-151

13. Molecular Monitoring of Chronic Myeloid Leukemia

    • Katherine Dominy, Katya Mokretar, Alistair G. Reid, Jamshid S. Khorashad

    Pages 153-173

14. Normalization in Human Glioma Tissue

    • Ana Paula Santin Bertoni, Isabele Cristiana Iser, Rafael Paschoal de Campos, Márcia Rosangela Wink

    Pages 175-190

15. qPCR Applications for the Determination of the Biological Age

    • Mauro Castagnetta, Ulrich Pfeffer, Aldo Chiesa, Elena Gennaro, Massimiliano Cecconi, Domenico Coviello et al.

    Pages 191-197

16. QuantStudio™ 12K Flex OpenArray® System as a Tool for High-Throughput Genotyping and Gene Expression Analysis

    • Chiara Broccanello, Letizia Gerace, Piergiorgio Stevanato

    Pages 199-208

17. Digital PCR and the QuantStudio™ 3D Digital PCR System

    • Marion Laig, Christie Fekete, Nivedita Majumdar

    Pages 209-231

18. Back Matter

    Pages 233-235

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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.

• Polymerase Chain Reaction
• RT-qPCR
• Primer design
• RT-PCR
• DNA sequencing
• qPCR

• Molecular Diagnostics, IRCCS, Istituto Giannina Gaslini, Genova, Italy

  Roberto Biassoni

• ASL3 Sistema Sanitario Regione Liguria, S.C. Laboratorio d’Analisi, Genova, Italy

  Alessandro Raso

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