RT-PCR Protocols

ISBN-13: 9781607616283 / Angielski / Twarda / 2010 / 356 str.

Nicola King

Opis

Once a tedious, highly skilled operation, reverse-transcription polymerase chain reaction (RT-PCR) has become a routine and invaluable technique used in most laboratories. In RT-PCR Protocols, Second Edition, expert researchers fully update the technologies presented in the popular previous edition, such as competitive RT-PCR, nested RT-PCR, RT-PCR from single cells, and RT-PCR for cloning. In addition, newer technologies are also explored, including multiplex RT-PCR, RT-LATE-PCR, and the greatly advanced field of real-time quantitative RT-PCR, while recent advances in creating the optimum RT-PCR reaction, e.g. RNA extraction, primer design, and reverse transcription, end the book with their indispensable input. Written in the highly successful Methods in Molecular Biology(TM) series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes sections, highlighting tips on troubleshooting and avoiding known pitfalls. User friendly and up-to-date, RT-PCR Protocols, Second Edition acts as a handy companion to scientists from numerous diverse backgrounds who wish to explore further the marvels of gene expression.

Recenzje

From the reviews of the second edition:
"The second edition of RT-PCR protocols and attempts to cover the most commonly used RT-PCR protocols, giving both beginners and experts a deeper understanding of the experimental set-up. ... There are often flow diagrams or pictures to aid understanding and the references are helpful, if you require additional information. ... a good investment for any lab interested in ensuring that their RT-PCR reactions are as productive and well-controlled as possible. ... Overall a useful and detailed text written in an informative and helpful style." (Hayley Evans, Immunology News, May, 2011)

Spis treści

Part I: The RT-PCR Detective: Hunting Down the Best Method 1. Single Cell RT-PCR on Mouse Embryos: A General Approach for Developmental Biology Wolfgang Mann and Thomas Haaf 2. Poly(A) cDNA Real-Time PCR for Indicator Gene Measurement in Cancer Melissa Oliveira-Cunha, Ajith K. Siriwardena, and Richard J. Byers 3. Transcriptome Profiling of Host-Microbe Interactions by Differential Display RT-PCR W. F. Leong and Vincent T. K. Chow 4. Quantitative RT-PCR Methods for Mature microRNA Expression Analysis Stephanie D. Fiedler, Martha Z. Carletti, and Lane K. Christenson 5. Detection of Influenza A Virus Neuraminidase and PB2 Gene Segments by One Step Reverse Transcription Polymerase Chain Reaction Alejandra Castillo Alvarez, Victoria Boyd, Richard Lai, Sandy Pineda, Hans G. Heine, and Ross Barnard 6. Detection and Identification of CD46 Splicing Isoforms by Nested RT-PCR Anita Szalmás, József Kónya, István Sziklai, and Tamás Karosi 7. Simultaneous Detection of Bluetongue Virus RNA, Internal Control GAPDH mRNA, and External Control Synthetic RNA by Multiplex Real-Time PCR Frank Vandenbussche, Elise Vandemeulebroucke, and Kris De Clercq 8. Detection of West Nile Viral RNA from Field-Collected Mosquitoes in Tropical Regions by Conventional and Real-Time RT-PCR Ana Silvia González-Reiche, María de Lourdes Monzón-Pineda, Barbara W. Johnson, and María Eugenia Morales-Betoulle 9. Detection of Antisense RNA Transcripts by Strand-Specific RT-PCR Eric C.H. Ho, Michael E. Donaldson, and Barry J. Saville 10. RT-PCR Amplification and Cloning of Large Viral Sequences Xiaofeng Fan and Adrian M. Di Bisceglie Part II: The RT-PCR Mathematician: Assessing Gene and RNA Expression 11. One-Step RT-LATE-PCR for mRNA and Viral RNADetection and Quantification Cristina Hartshorn and Lawrence J. Wangh 12. Changes in Gene Expression of Caveolin-1 after Inflammatory Pain Using Quantitative Real-Time PCR Fiza Rashid-Doubell 13. Real-Time Quantitative Reverse Transcriptase Polymerase Chain Reaction Hongxin Fan and Ryan S. Robetorye 14. The Use of Comparative Quantitative RT-PCR to Investigate the Effect of Cysteine Incubation on GPx1 Expression in Freshly Isolated Cardiomyocytes Nicola King 15. The Renaissance of Competitive PCR as an Accurate Tool for Precise Nucleic Acid Quantification Lorena Zentilin and Mauro Giacca Part III: The RT-PCR Master Chef: Finding the Right Ingredients 16. Skeletal Muscle RNA Extraction in Preparation for RT-PCR Janelle P. Mollica 17. Reverse Transcription of the Ribonucleic Acid: A First Step in RT-PCR Assay Fadia Haddad and Kenneth M. Baldwin 18. Primer Design for RT-PCR Kelvin Li and Anushka Brownley 19. Hot Start PCR Natasha Paul, Jonathan Shum, and Tony Le 20. Real-time RT-PCR for Automated Detection of HIV-1 RNA During Blood Donor Screening Jens Müller

Blurb

Once a tedious, highly skilled operation, reverse-transcription polymerase chain reaction (RT-PCR) has become a routine and invaluable technique used in most laboratories. In RT-PCR Protocols, Second Edition, expert researchers fully update the technologies presented in the popular previous edition, such as competitive RT-PCR, nested RT-PCR, RT-PCR from single cells, and RT-PCR for cloning. In addition, newer technologies are also explored, including multiplex RT-PCR, RT-LATE-PCR, and the greatly advanced field of real-time quantitative RT-PCR, while recent advances in creating the optimum RT-PCR reaction, e.g. RNA extraction, primer design, and reverse transcription, end the book with their indispensable input. Written in the highly successful Methods in Molecular Biology™ series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes sections, highlighting tips on troubleshooting and avoiding known pitfalls.

User friendly and up-to-date, RT-PCR Protocols, Second Edition acts as a handy companion to scientists from numerous diverse backgrounds who wish to explore further the marvels of gene expression.