RT-PCR Protocols Second Edition

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.

The RT-PCR Detective: Hunting Down the Best Method.- Single Cell RT-PCR on Mouse Embryos: A General Approach for Developmental Biology.- Poly(A) cDNA Real-Time PCR for Indicator Gene Measurement in Cancer.- Transcriptome Profiling of Host–Microbe Interactions by Differential Display RT-PCR.- Quantitative RT-PCR Methods for Mature microRNA Expression Analysis.- Detection of Influenza A Virus Neuraminidase and PB2 Gene Segments by One Step Reverse Transcription Polymerase Chain Reaction.- Detection and Identification of CD46 Splicing Isoforms by Nested RT-PCR.- Simultaneous Detection of Bluetongue Virus RNA, Internal Control GAPDH mRNA, and External Control Synthetic RNA by Multiplex Real-Time PCR.- Detection of West Nile Viral RNA from Field-Collected Mosquitoes in Tropical Regions by Conventional and Real-Time RT-PCR.- Detection of Antisense RNA Transcripts by Strand-Specific RT-PCR.- RT-PCR Amplification and Cloning of Large Viral Sequences.- The RT-PCR Mathematician: Assessing Gene and RNA Expression.- One-Step RT-LATE-PCR for mRNA and Viral RNA Detection and Quantification.- Changes in Gene Expression of Caveolin-1 After Inflammatory Pain Using Quantitative Real-Time PCR.- Real-Time Quantitative Reverse Transcriptase Polymerase Chain Reaction.- The Use of Comparative Quantitative RT-PCR to Investigate the Effect of Cysteine Incubation on GPx1 Expression in Freshly Isolated Cardiomyocytes.- The Renaissance of Competitive PCR as an Accurate Tool for Precise Nucleic Acid Quantification.- The RT-PCR Master Chef: Finding the Right Ingredients.- Skeletal Muscle RNA Extraction in Preparation for RT-PCR.- Reverse Transcription of the Ribonucleic Acid: The First Step in RT-PCR Assay.- Primer Design for RT-PCR.- Hot Start PCR.- Real-time RT-PCR for Automated Detection ofHIV-1 RNA During Blood Donor Screening.