1st Edition

PCR/RT- PCR in situ
Light and Electron Microscopy




ISBN 9780849300417
Published September 27, 2002 by CRC Press
432 Pages 21 Color & 237 B/W Illustrations

USD $250.00

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Book Description

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.

Table of Contents

GENERAL PRINCIPLES
PREPARATION OF SAMPLES
PRETREATMENTS
REVERSE TRANSCRIPTION (RT)
POLYMERASE CHAIN REACTION (PCR)
HYBRIDIZATION
REVELATION
ELECTRON MICROSCOPY
CONTROLS AND PROBLEMS
TYPICAL PROTOCOLS
EXAMPLES OF OBSERVATIONS
APPENDICES
A-Equipment
B-Reagents
GLOSSARY
INDEX

Short TOC
General Principles. Preparation of Samples. Pretreatments. Reverse Transcription (RT). Polymerase Chain Reaction (PCR). Hybridization. Revelation. Electron Microscopy. Controls and Problems. Typical Protocols. Examples of Observations. Appendices. Glossary. Index

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