Protocols for Nucleic Acid Analysis by Nonradioactive Probes

ISBN-13: 9781617375644 / Angielski / Miękka / 2010 / 321 str.

Elena Hilario; John F. MacKay

Opis

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.
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Recenzje

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)

Spis treści

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