Bartlett John M.S., Stirling David (eds.) PCR Protocols

2nd edition. — Humana Press, 2003. — 519 p. — (Methods in Molecular Biology 226) — ISBN: 978-0-89603-627-7Drawing on the proven qualities of the much praised and widely used first edition, John M. S. Bartlett and David Stirling have thoroughly updated and dramatically expanded the number of protocols to take advantage of the newest technologies used in all branches of research and clinical medicine today. These successful methods include real-time PCR, SNP analysis, nested PCR, direct PCR, and long-range PCR. Among the highlights are chapters on genome profiling by SAGE, differential display and chip technologies, the amplification of whole genome DNA by random degenerate oligonucleotide PCR, and the refinement of PCR methods for the analysis of fragmented DNA from fixed tissues. In situ PCR methods and their application in parallel with other methods, such as immunohistochemistry, are also included. Each fully tested protocol is described in step-by-step detail by an established expert in the field and includes a background introduction outlining the principle behind the technique, equipment and reagent lists, tips on troubleshooting and avoiding known pitfalls, and, where needed, a discussion of the interpretation and use of results.Cutting-edge and highly practical, PCR Protocols, Second Edition provides both novice and experienced investigators with an up-to-date compendium of powerful PCR methods for easy reference and consultation in the day-to-day performance of PCR-based experimentation, one that will enhance understanding of PCR, satisfy current needs, and point to powerful future applications.A Short History of the Polymerase Chain Reaction
PCR Patent Issues
Equipping and Establishing a PCR Laboratory
Quality Control in PCR
Extraction of Nucleic Acid Templates
Extraction of DNA from Whole Blood
DNA Extraction from Tissue
Extraction of DNA from Microdissected Archival Tissues
RNA Extraction from Blood
RNA Extraction from Frozen Tissue
RNA Extraction from Tissue Sections
Dual DNA/RNA Extraction
DNA Extraction from Fungi, Yeast, and Bacteria
Isolation of RNA Viruses from Biological Materials
Extraction of Ancient DNA
DNA Extraction from Plasma and Serum
Technical Notes for the Detection of Nucleic Acids
Technical Notes for the Recovery and Purification of PCR Products from Acrylamide Gels
PCR Primer Design
Optimization of Polymerase Chain Reactions
Subcycling PCR for Long-Distance Amplifications of Regions with High and Low Guanine-Cystine Content
Rapid Amplification of cDNA Ends
Randomly Amplified Polymorphic DNA Fingerprinting
Microsphere-Based Single Nucleotide Polymorphism Genotyping
Ligase Chain Reaction
Nested RT-PCR in a Single Closed Tube
Direct PCR from Serum
Long PCR Amplification of Large Fragments of Viral Genomes
Long PCR Methodology
Qualitative and Quantitative PCR
Ultrasensitive PCR Detection of Tumor Cells in Myeloma
Ultrasensitive Quantitative PCR to Detect RNA Viruses
Quantitative PCR for cAMP RI Alpha mRNA
Quantitation of Multiple RNA Species