Protocols for nucleic acid analysis by nonradioactive probes

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(tm) 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.

Duckchul ParkElspeth MacRaeElena HilarioLisa S. Gellen and Glenn M. Wall-Manning and Chris H. SissonsChristoph Peterhaensel and Dagmar Weier and Thomas LahayeElena Hilario and Tiffany F. Bennell and Erik RikkerinkSaeed A. Khan and Mohamed S. NawazShigeharu Moriya and Satoko Noda and Moriya Ohkuma and Toshiaki KudoYukinori EguchiDarren J. Day and Eli M. Mrkusich and John H. MillerMaria Rosa Ponce and Victor Quesada and Andrea Hricova and Jose Luis MicolAnnelie Pernthaler and Jakob PernthalerJohn MackaySukru Tuzmen and Jeff Kiefer and Spyro MoussesJasmien Hoebeeck and Frank Speleman and Jo VandesompeleHarald H. KesslerJohn Mackay and Olfert LandtNorman H. Lee and Alexander I. SaeedGaspar Taroncber-Oldenburg and Bess B. Ward

Preface	p. v
Contributors	p. ix
Part I Nucleic Acid Extractions
1 Genomic DNA Isolation From Different Biological Materials	p. 3
2 Extraction of Plant RNA	p. 15
Part II Indirect Detection
3 Overview of Hybridization and Detection Techniques	p. 27
4 Checkerboard DNA-DNA Hybridization Technology Using Digoxigenin Detection	p. 39
5 Nonradioactive Northern and Southern Analyses From Plant Samples	p. 69
6 Screening a BAC Library With Nonradioactive Overlapping Oligonucleotide (Overgo) Probes	p. 79
7 Direct In-Gel Hybridization of DNA With Digoxigenin-Labeled Probes	p. 93
8 In Situ Hybridization of Termite Microbes	p. 105
Part III Fluorescent Labeling and Detection
9 RNA Electrophoretic Mobility Shift Assay Using a Fluorescent DNA Sequencer	p. 115
10 Comparative Quantitation of mRNA Expression in the Central Nervous System Using Fluorescence In Situ Hybridization	p. 125
11 Visualization of Gene Expression by Fluorescent Multiplex Reverse Transcriptase-PCR Amplification	p. 143
12 Fluorescence In Situ Hybridization for the Identification of Environmental Microbes	p. 153
Part IV Kinetic ("Real-Time") PCR
13 Introduction to Kinetic (Real-Time) PCR	p. 167
14 Validation of Short Interfering RNA Knockdowns by Quantitative Real-Time-PCR	p. 177
15 Real-Time Quantitative PCR as an Alternative to Southern Blot or Fluorescence In Situ Hybridization for Detection of Gene Copy Number Changes	p. 205
16 Design and Work-Up of a New Molecular Diagnostic Assay Based on Real-Time PCR	p. 227
17 Real-Time PCR Fluorescent Chemistries	p. 237
Part V Microarrays
18 Microarrays: An Overview	p. 265
19 Oligonucleotide Microarrays for the Study of Coastal Microbial Communities	p. 301
Index	p. 317

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