PCR Protocols

• 1 A Short History of the Polymerase Chain Reaction.- 2 PCR Patent Issues.- 3 Equipping and Establishing a PCR Laboratory.- 4 Quality Control in PCR.- 5 Extraction of Nucleic Acid Templates.- 6 Extraction of DNA from Whole Blood.- 7 DNA Extraction from Tissue.- 8 Extraction of DNA from Microdissected Archival Tissues.- 9 RNA Extraction from Blood.- 10 RNA Extraction from Frozen Tissue.- 11 RNA Extraction from Tissue Sections.- 12 Dual DNA/RNA Extraction.- 13 DNA Extraction from Fungi, Yeast, and Bacteria.- 14 Isolation of RNA Viruses from Biological Materials.- 15 Extraction of Ancient DNA.- 16 DNA Extraction from Plasma and Serum.- 17 Technical Notes for the Detection of Nucleic Acids.- 18 Technical Notes for the Recovery and Purification of PCR Products from Acrylamide Gels.- 19 PCR Primer Design.- 20 Optimization of Polymerase Chain Reactions.- 21 Subcycling PCR for Long-Distance Amplifications of Regions with High and Low Guanine-Cystine Content: Amplification of the Intron 22 Inversion of the FVIII Gene.- 22 Rapid Amplification of cDNA Ends.- 23 Randomly Amplified Polymorphic DNA Fingerprinting: The Basics.- 24 Microsphere-Based Single Nucleotide Polymorphism Genotyping.- 25 Ligase Chain Reaction.- 26 Nested RT-PCR in a Single Closed Tube.- 27 Direct PCR from Serum: Application to Viral Genome Detection.- 28 Long PCR Amplification of Large Fragments of Viral Genomes: A Technical Overview.- 29 Long PCR Methodology.- 30 Qualitative and Quantitative PCR: A Technical Overview.- 31 Ultrasensitive PCR Detection of Tumor Cells in Myeloma.- 32 Ultrasensitive Quantitative PCR to Detect RNA Viruses.- 33 Quantitative PCR for cAMP RI Alpha mRNA: Use of Site-Directed Mutation and PCR Mimics.- 34 Quantitation of Multiple RNA Species.- 35 Differential Display: A Technical Overview.- 36 AU-Differential Display, Reproducibility of a Differential mRNA Display Targeted to AU Motifs.- 37 PCR Fluorescence Differential Display.- 38 Microarray Analysis Using RNA Arbitrarily Primed PCR.- 39 Oligonucleotide Arrays for Genotyping: Enzymatic Methods for Typing Single Nucleotide Polymorphisms and Short Tandem Repeats.- 40 Serial Analysis of Gene Expression.- 41 Mutation and Polymorphism Detection: A Technical Overview.- 42 Combining Multiplex and Touchdown PCR for Microsatellite Analysis.- 43 Detection of Microsatellite Instability and Loss of Heterozygosity Using DNA Extracted from Formalin-Fixed Paraffin-Embedded Tumor Material by Fluorescence-Based Multiplex Microsatellite PCR.- 44 Reaction of Shadow Band Synthesis During PCR Amplification of Repetitive Sequences from Modern and Ancient DNA.- 45 Degenerate Oligonucleotide-Primed PCR.- 46 Mutation Detection Using RT-PCR-RFLP.- 47 Multiplex Amplification Refractory Mutation System for the Detection of Prothrombotic Polymorphisms.- 48 PCR-SSCP Analysis of Polymorphism: A Simple and Sensitive Method for Detecting Differences Between Short Segments of DNA.- 49 Sequencing: A Technical Overview.- 50 Preparation and Direct Automated Cycle Sequencing of PCR Products.- 51 Nonradioactive PCR Sequencing Using Digoxigenin.- 52 Direct Sequencing by Thermal Asymmetric PCR.- 53 Analysis of Nucleotide Sequence Variations by Solid-Phase Minisequencing.- 54 Direct Sequencing with Highly Degenerate and Inosine-Containing Primers.- 55 Determination of Unknown Genomic Sequences Without Cloning.- 56 Cloning PCR Products for Sequencing in M13 Vectors.- 57 DNA Rescue by the Vectorette Method.- 58 Technical Notes for Sequencing Difficult Templates.- 59 PCR-Based Detection of Nucleic Acids in Chromosomes, Cells, and Tissues: Technical Considerations on PRINS and In Situ PCR, and Comparison with In Situ Hybridization.- 60 Cycling Primed In Situ Amplification.- 61 Direct and Indirect In Situ PCR.- 62 Reverse Transcriptase In Situ PCR: New Methods in Cellular Interrogation.- 63 Primed In Situ Nucleic Acid Labeling Combined with Immunocytochemistry to Simultaneously Localize DNA and Proteins in Cells and Chromosomes.- 64 Cloning and Mutagenesis: A Technical Overview.- 65 Using T4 DNA Polymerase to Generate Clonable PCR Products.- 66 A T-Linker Strategy for Modification and Directional Cloning of PCR Products.- 67 Cloning Gene Family Members Using PCR with Degenerate Oligonucleotide Primers.- 68 cDNA Libraries from a Low Amount of Cells.- 69 Creation of Chimeric Junctions, Deletions, and Insertions by PCR.- 70 Recombination and Site-Directed Mutagenesis Using Recombination PCR.- 71 Megaprimer PCR: Application in Mutagenesis and Gene Fusion.

Reviews of the First Edition: "...succeeds in covering a wider range of more general applicable methods than...its predecessors ...provid[es] an extensive range of versatile, expedient, and readily applicable PCR protocols."-Trends in Cell Biology From reviews of the first edition... "...useful in virtually all disciplines of the life sciences...a pleasure [to] read and use..." -American Journal of Physiology: Lung Cellular and Molecular Physiology "...an excellent resource...has potential value for anyone looking for new ways to use PCR or new tricks to make their PCR work better." -Biopharm "...provides an extensive range of versatile, expedient, and readily applicable PCR protocols." -Trends in Cell Biology