PCR/RT- PCR in situ: Light and Electron Microscopy / Edition 1

by Gerard Morel, Mireille Raccurt

Overview

Although the polymerase chain reaction has revolutionized genetic analysis by amplifying rare nucleic acid sequences, the in situ application is the only method that allows the localization of amplified signal within tissue structure. The applications of in situ polymerase chain reaction have greatly enhanced the field of investigation in many disciplines, including viral infections, gene modification, tumor diagnosis, gene therapy, and cellular distribution of rare mRNA copies.

PCR/RT-PCR in situ: Light and Electron Microscopy covers methods of in situ polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR), two new approaches in visualizing very low amounts of DNA and RNA in tissues and cell cultures at the light and electron microscopy levels. Written by experts in this field, the book provides theoretical consideration, as well as practical approaches to in situ PCR. The authors provide detailed protocols for each step, including the preparation of tissue samples, the rationale for the design of primers and revelation. They also emphasize the need for appropriate controls to meet the requirements of in situ PCR and RT-PCR specificity. Organized in a user-friendly two-column format, this book will provide you with tools necessary to perform and optimize these sensitive and powerful techniques in your research protocols.

Product Details

ISBN-13:	9780849300417
Publisher:	Taylor & Francis
Publication date:	09/27/2002
Series:	Methods in Visualization , #6
Edition description:	New Edition
Pages:	444
Product dimensions:	6.80(w) x 10.00(h) x 1.20(d)

About the Author

Gérard Morel, Ph.D., D.Sc., is a research director at the National Center of Scientific Research (CNRS), at University Claude Bernard-Lyon 1, Villeurbanne, France. Mireille Raccurt, Ph.D., is an engineer in biology in a laboratory of the CNRS (Molecular Physiology), at Claude Bernard-Lyon 1 University, Villeurbanne, France.

Table of Contents

General Introduction Chapter 1 - General Principles Chapter 2 - Preparation of Samples Chapter 3 - Pretreatments Chapter 4 - Reverse Transcription (RT) Chapter 5 - Polymerase Chain Reation (PCR) Chapter 6 - Hybridization Chapter 7 - Revelation Chapter 8 - Electron Microscopy Chapter 9 - Controls and Problems Chapter 10 - Typical Protocols Chapter 11 - Examples of Observations