Quantitative Real-Time PCR Methods and Protocols

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. 

1. A Quarter Century and the PCR Applied Techniques and the Fields of Use Are Still Increasing in Numbers

            Alessandro Raso and Roberto Biassoni

 

2. Parameters for Successful PCR Primer Design

            Stephen A. Bustin, Reinhold Mueller, and Tania Nolan

 

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

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

 

4. Enhanced Probe-Based RT-qPCR Quantification of MicroRNAs Using Poly(A)tailing and 5' Adaptor Ligation

            Valentin Vautrot and Isabelle Behm-Ansmant

 

5. A Novel System to Discriminate HLA-C mir148a Binding Site by Allele-Specific Quantitative PCR

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

 

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

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

 

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

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

 

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

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

 

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

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

 

10. Gene Expression Analysis in Bacteria by RT-qPCR

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

 

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

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

 

12. Molecular Monitoring of Chronic Myeloid Leukemia

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

 

13. Normalization in Human Gliomas Tissue

            Ana Paula Santin Bertoni, Isabele Cristina Iser Marson, Rafael Paschoal de Campos, and Márcia Rosangela Wink

 

14. qPCR Applications for the Determination of the Biological Age

            Mauro Castagnetta, Ulrich Pfeffer, Aldo Chiesa, Elena Gennaro, Massimiliano Cecconi, Domenico Coviello, and Nicoletta Sacchi

 

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

            Chiara Broccanello, Letizia Gerace, and Piergiorgio Stevanato

 

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

            Marion Laig, Christie Fekete, and Nivedita Majumdar