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Protocols for Nucleic Acid Analysis by Nonradioactive Probes

Series: Methods in Molecular Biology

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by Elena Hilario and John F. MacKay

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Protocols for Nucleic Acid Analysis by Nonradioactive Probes

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Main description:

Protocols for Nucleic Acid Analysis by Non-radioactive Probes, Second Edition provides a firm background on the basic preparative protocols required for the analysis of nucleic acids by nonradioactive methods. Presenting the methodologies using amazing new applications, this volume offers guide chapters on nucleic acid extractions, preparation of nucleic acid blots, and labeling of nucleic acids with nonradioactive haptens. New fluorescent techniques such as Real Time PCR and microarrays are also included, allowing users to get a nonradioactive protocol implemented in the laboratory with minimum adaptation required and fastest time to results.

The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

Feature:

Readily reproducible protocols for the analysis of nucleic acids by nonradioactive methods
New fluorescent techniques such as Real Time PCR and microarrays
Follow step-by-step instructions that ensure successful results
Master tricks of the trade, troubleshoot, and avoid known pitfalls

Contents:

Contents
(NOTE: Chapters not in MiMB/MiMM format are highlighted)
Preface
Contributors

A. Nucleic Acid Extractions
1. Genomic DNA Isolation from Different Biological Materials
Duckchul Park
2 Extraction of Plant RNA
Elspeth MacRae

B. Indirect Detection
3 Overview on Hybridization and Detection Techniques
Elena Hilario
4 Checkerboard DNA:DNA Hybridization Technology using DIG Detection
Glenn Wall-Manning, Lisa S. Gellen, and Chris H. Sissons
5 Non-radioactive Northern and Southern analyses from Plant Samples
Christoph Peterhaensel, Dagmar Weier, and Thomas Lahaye
6 Screening a BAC library with Non-Radioactive Overlapping Oligonucleotide (Overgo) Probes
Elena Hilario, Tiffany Bennell, and Erik Rikkerink
7 Direct In-gel Hybridization of DNA with Digoxigenin-Labeled Probes
Saeed A. Khan and Mohamed S. Nawaz
8 In situ Hybridization of Termite Microbes
Shigeharu Moriya , Satoko Noda, Moriya Ohkuma and Toshiaki Kudo

C. Fluorescent Labeling and Detection
9 RNA Electrophoretic Mobility Shift Assay using a Fluorescent DNA Sequencer
Yukinori Eguchi
10 Comparative Quantitation of mRNA Expression in the Central Nervous System using Fluorescence In situ Hybridization
Darren Day, Eli M. Mrkusich, and John H. Miller
11 Visualization of Gene Expression by Fluorescent Multiplex RT-PCR Amplification
María Rosa Ponce, Víctor Quesada, Andrea Hricová, and José Luis Micol
12 Fluorescence In situ Hybridization for the Identification of Environmental Microbes
Annelie Pernthaler and Jakob Pernthaler

D. Kinetic ('Real-Time') PCR
13 Introduction to Kinetic (Real-Time) PCR
John Mackay
14 Validation of siRNA Knockdowns by Quantitative Real-Time PCR
Sukru Tuzmen
15 Real-Time Quantitative PCR as an Alternative to Southern Blot or FISH for Detection of Gene-Copy NumberChanges
Jasmien Hoebeeck, Frank Speleman, and Jo Vandesompele
16 Design and Working up a New Molecular Diagnostic Assay Based on Real-Time PCR
Harald Kessler
17 Real-Time PCR Fluorescent Chemistries
John Mackay and Olfert Landt

E. Microarrays
18 Microarrays: An Overview
Norman H. Lee and Alexander I. Saeed
19 Oligonucleotide Microarrays for the Study of Coastal Microbial Communities
Gaspar Taroncher-Oldenburg and Bess B. Ward

PRODUCT DETAILS

ISBN-13: 9781588294302
Publisher: Springer (Humana Press)
Publication date: January, 2007
Pages: 321
Weight: 721g
Availability: Not available (reason unspecified)
Subcategories: Biochemistry, Genetics

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CUSTOMER REVIEWS

Average Rating

From the reviews of the second edition:

...this book provides a collection of good protocols which are in general clearly written. They can be easily employed on the bench without being a specialist in a particular fields. - FEBS Letters

...Covers the basics of a variety of probe-labelling systems very well...a very interesting read. It could save you money and trouble from the radiation safety officer.-Biotechnology and Applied Biochemistry

"This is a set of protocols for various types of nucleic acid analysis using nonradiocative methods. … The goal is to provide protocols for nucleic acid analysis using other than radioactive reporter systems. This approach is commonplace at present, so a volume of practical utility has merit. … The target audience would be research laboratory personnel working in this general area. The editors have recruited a group of contributors with worldwide representation." (Eugene A Davidson, Doody’s Review Service, November, 2007)

"The editors of this volume have gathered together a variety of protocols for anyone who wishes to use nonradioactive methods for the analysis of their chosen nucleic acid molecules. ... Combined with the excellent overviews on microarrays as well as hybridization and detection techniques, and several useful assays for the application of these techniques for nucleic acid analysis, scientists will find this book handy for their research and educators will adopt it as a useful teaching tool for advanced students." (Tzvi Tzfira, The Quarterly Review of Biology, Vol. 83, 2008)