PCR/RT- PCR in situ: Light and Electron Microscopy, 1st Edition (Hardback) book cover

PCR/RT- PCR in situ

Light and Electron Microscopy, 1st Edition

By Gerard Morel, Mireille Raccurt

CRC Press

432 pages | 21 Color Illus. | 237 B/W Illus.

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pub: 2002-09-27
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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

About the Series

Methods in Visualization

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Subject Categories

BISAC Subject Codes/Headings:
SCI017000
SCIENCE / Life Sciences / Cytology
SCI047000
SCIENCE / Microscopes & Microscopy
SCI049000
SCIENCE / Life Sciences / Biology / Molecular Biology
SCI086000
SCIENCE / Life Sciences / General