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™ 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.

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 Includes supplementary material: sn.pub/extras

Inhalt
Nucleic Acid Extractions.- Genomic DNA Isolation From Different Biological Materials.- Extraction of Plant RNA.- Indirect Detection.- Overview of Hybridization and Detection Techniques.- Checkerboard DNA-DNA Hybridization Technology Using Digoxigenin Detection.- Nonradioactive Northern and Southern Analyses From Plant Samples.- Screening a BAC Library With Nonradioactive Overlapping Oligonucleotide (Overgo) Probes.- Direct In-Gel Hybridization of DNA With Digoxigenin-Labeled Probes.- In Situ Hybridization of Termite Microbes.- Fluorescent Labeling and Detection.- RNA Electrophoretic Mobility Shift Assay Using a Fluorescent DNA Sequencer.- Comparative Quantitation of mRNA Expression in the Central Nervous System Using Fluorescence In Situ Hybridization.- Visualization of Gene Expression by Fluorescent Multiplex Reverse Transcriptase-PCR Amplification.- Fluorescence In Situ Hybridization for the Identification of Environmental Microbes.- Kinetic (Real-Time) PCR.- to Kinetic (Real-Time) PCR.- Validation of Short Interfering RNA Knockdowns by Quantitative Real-Time PCR.- Real-Time Quantitative PCR as an Alternative to Southern Blot or Fluorescence In Situ Hybridization for Detection of Gene Copy Number Changes.- Design and Work-Up of a New Molecular Diagnostic Assay Based on Real-Time PCR.- Real-Time PCR Fluorescent Chemistries.- Microarrays.- Microarrays.- Oligonucleotide Microarrays for the Study of Coastal Microbial Communities.