About the Book
A panel of highly regarded molecular biologists and clinical researchers describe in detail their most novel, useful, and interesting RT-PCR applications. Here the newcomer will find readily reproducible protocols for highly sensitive detection and quantification of gene expression, the in situ localization of gene expression in tissue, and the cloning of genes, as well as for analyzing T-cell clones and the differential expression of genes. For the expert seeking to extend the usefulness of RT-PCR, there are user-friendly applications that complement the latest technological advances, including laser-capture microdissection (LCM), real-time and quantitative PCR, microarray technology, cDNA cloning, and antibody engineering.
Table of Contents:
Part I. Introduction RT-PCR in Biomedicine: Opportunities Arising from the New Accessibility of mRNA Joe O'Connell The Basics of RT-PCR: Some Practical Considerations Joe O'Connell Part II. Highly Sensitive Detection and Analysis of mRNA Description of Quantitative Competitive RT-PCR Technique to Analyze Minute Amounts of Different mRNAs in Small Tissue Samples Susanne Greber-Platzer, Brigitte Balcz, Christine Fleischmann, and Gert Lubec Detection of mRNA Expression and Alternative Splicing in a Single Cell Tsutomu Kumazaki Nested RT-PCR: Sensitivity Controls are Essential to Determine the Biological Significance of Detected mRNA Triona Goode, Wen-Zhe Ho, Terry O'Connor, Sandra Busteed, Steven D. Douglas, Fergus Shanahan, and Joe O'Connell Part III. Quantitative RT-PCR Quantitative RT-PCR: A Review of Current Methodologies Caroline Joyce Rapid Development of a Quantitative-Competitive (QC) RT-PCR Assay Using a Composite Primer Approach Joe O'Connell, Aileen Houston, Raymond Kelly, Darren O'Brien, Aideen Ryan, Michael W. Bennett, and Kenneth Nally Quantitation of Gene Expression by RT-PCR and HPLC Analysis of PCR Products Franz Bachmair, Christian G. Huber, and Guenter Daxenbichler Time-Resolved Fluorometric Detection of Cytokine mRNAs Amplified by RT-PCR Kaisa Nieminen, Markus Halminen, Matti Waris, Mika Makela, Johannes Savolainen, Minna Sjoroos, and Jorma Ilonen Mimic-Based RT-PCR Quantitation of Substance P mRNA in Human Mononuclear Phagocytes and Lymphocytes Jian-Ping Lai, Steven D. Douglas, and Wen-Zhe Ho Part IV. Detection and Analysis of RNA Viruses Detection and Quantification of the Hepatitis C Viral Genome Liam J. Fanning Semi-Quantitative Detection of Hepatitis C Virus RNA by "Real-Time" RT-PCR Joerg F. Schlaak RT-PCR for the Assessment of Genetically Heterogenous Populations of the Hepatitis C Virus Brian Mullan, Liam Fanning, Fergus Shanahan, and Daniel G. Sullivan Part V. In Situ Localization of mRNA Expression In Situ Immuno-PCR: A Newly Developed Method for Highly Sensitive Antigen Detection In Situ Yi Cao RT-PCR from Laser Capture Microdissected Samples Tatjana Crnogorac-Jurcevic, Torsten O. Nielsen, and Nick R. Lemoine Mycobacterium Paratuberculosis Detected by Nested PCR in Intestinal Granulomas Isolated by Laser Capture Microdissection in Cases of Crohn's Disease Paul Ryan, Simon Aarons, Michael W. Bennett, Gary Lee, Gerald C. O'Sullivan, Joe O'Connell, and Fergus Shanahan RT-PCR-Based Approaches to Generate Probes for mRNA Detection by In Situ Hybridization Joe O'Connell Part VI. Differential mRNA Expression Amplified RNA for Gene Array Hybridizations Valentina I. Shustova and Stephen J. Meltzer Semiquantitative Determination of Differential Gene Expression in Primary Tumors and Matched Metastases by RT-PCR: Comparison with Other Methods Benno Mann and Christoph Hanski Part VII. Genetic Analysis Detection of Single Nucleotide Polymorphisms Using a Non-Isotopic RNase Cleavage Assay Frank Waldron-Lynch, Claire Adams, Michael G. Molloy, and Fergal O'Gara Part VIII. RT-PCR in Immunology Detection of Clonally Expanded T Cells by RT-PCR-SSCP and Nucleotide Sequencing of T Cell Receptor b-CDR3 Regions Manae Suzuki Kurokawa, Kusuki Nishioka, and Tomohiro Kato Generation of scFv from a Phage Display Mini-Library Derived from Tumor-Infiltrating B Cells Nadege Gruel, Beatrix Kotlan, Marie Beuzard, and Jean-Luc Teillaud Generation of Murine scFv Intrabodies from B Cell Hybridomas Chang Hoon Nam, Sandrine Moutel, and Jean-Luc Teillaud Quantitation of mRNA Levels by RT-PCR in Cells Purified by FACS: Application to Peripheral Cannabinoid Receptors in Leukocyte Subsets Jean Marchand and Pierre Carayon Part IX. RT-PCR in Antisense Technology Detection of Antisense RNA Transcripts by Antisense RT-PCR Michael C. Yeung and Allan S. Lau Part X. RT-PCR in cDNA Cloning RT-PCR in cDNA Library Construction Vincent Healy An RT-PCR-Based Protocol for the Rapid Generation of Large, Representative cDNA Libraries Suitable for Expression Screening Joe O'Connell Index
Review :
...RT-PCR succeeds in its goal to provide the reader with useful applications relevant to the wider research community. - Clinical Chemistry It is clearly written and can be easily understood by the readers. It is recommended for students, geneticists, immunologists and other specialists in biomedicine. - Folia Microbiologica
