RT-PCR Protocols

by King, Nicola

• ISBN13: 9781607616283
• ISBN10: 1607616289
• Edition: 2nd
• Format: Hardcover
• Copyright: 2010-04-21
• Publisher: Humana Pr Inc

Summary

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.

Table of Contents

Preface	p. v
Contributors	p. ix
The RT-PCR Detective: Hunting Down the Best Method
Single Cell RT-PCR on Mouse Embryos: A General Approach for Developmental Biology	p. 3
Poly(A) cDNA Real-Time PCR for Indicator Gene Measurement in Cancer	p. 13
Transcriptome Profiling of Host-Microbe Interactions by Differential Display RT-PCR	p. 33
Quantitative RT-PCR Methods for Mature microRNA Expression Analysis	p. 49
Detection of Influenza A Virus Neuraminidase and PB2 Gene Segments by One Step Reverse Transcription Polymerase Chain Reaction	p. 65
Detection and Identification of CD46 Splicing Isoforms by Nested RT-PCR	p. 83
Simultaneous Detection of Bluetongue Virus RNA, Internal Control GAPDH mRNA, and External Control Synthetic RNA by Multiplex Real-Time PCR	p. 97
Detection of West Nile Viral RNA from Field-Collected Mosquitoes in Tropical Regions by Conventional and Real-Time RT-PCR	p. 109
Detection of Antisense RNA Transcripts by Strand-Specific RT-PCR	p. 125
RT-PCR Amplification and Cloning of Large Viral Sequences	p. 139
The RT-PCR Mathematician: Assessing Gene and RNA Expression
One-Step RT-LATE-PCR for mRNA and Viral RNA Detection and Quantification	p. 153
Changes in Gene Expression of Caveolin-l After Inflammatory Pain Using Quantitative Real-Time PCR	p. 187
Real-Time Quantitative Reverse Transcriptase Polymerase Chain Reaction	p. 199
The Use of Comparative Quantitative RT-PCR to Investigate the Effect of Cysteine Incubation on GPxl Expression in Freshly Isolated Cardiomyocytes	p. 215
The Renaissance of Competitive PCR as an Accurate Tool for Precise Nucleic Acid Quantification	p. 233
The RT-PCR Master Chef: Finding the Right Ingredients
Skeletal Muscle RNA Extraction in Preparation for RT-PCR	p. 251
Reverse Transcription of the Ribonucleic Acid: The First Step in RT-PCR Assay	p. 261
Primer Design for RT-PCR	p. 271
Hot Start PCR	p. 301
Real-time RT-PCR for Automated Detection of HIV-1 RNA During Blood Donor Screening	p. 319
Index	p. 337
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