| Preface | p. v |
| Contributors | p. xiii |
| Introduction to PCR | |
| A Short History of the Polymerase Chain Reaction | p. 3 |
| PCR Patent Issues | p. 7 |
| Equipping and Establishing a PCR Laboratory | p. 15 |
| Quality Control in PCR | p. 21 |
| Preparation of Nucleic Acid Templates | |
| Extraction of Nucleic Acid Templates | p. 27 |
| Extraction of DNA from Whole Blood | p. 29 |
| DNA Extraction from Tissue | p. 33 |
| Extraction of DNA from Microdissected Archival Tissues | p. 35 |
| RNA Extraction from Blood | p. 43 |
| RNA Extraction from Frozen Tissue | p. 45 |
| RNA Extraction from Tissue Sections | p. 47 |
| Dual DNA/RNA Extraction | p. 49 |
| DNA Extraction from Fungi, Yeast, and Bacteria | p. 53 |
| Isolation of RNA Viruses from Biological Materials | p. 55 |
| Extraction of Ancient DNA | p. 57 |
| DNA Extraction from Plasma and Serum | p. 63 |
| Technical Notes for the Detection of Nucleic Acids | p. 65 |
| Technical Notes for the Recovery and Purification of PCR Products from Acrylamide Gels | p. 77 |
| Basic PCR Methods | |
| PCR Primer Design | p. 81 |
| Optimization of Polymerase Chain Reactions | p. 89 |
| 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 | p. 101 |
| Rapid Amplification of cDNA Ends | p. 105 |
| Randomly Amplified Polymorphic DNA Fingerprinting: The Basics | p. 117 |
| Microsphere-Based Single Nucleotide Polymorphism Genotyping | p. 123 |
| Ligase Chain Reaction | p. 135 |
| Nested RT-PCR in a Single Closed Tube | p. 151 |
| Direct PCR from Serum: Application to Viral Genome Detection | p. 161 |
| Long PCR Amplification of Large Fragments of Viral Genomes: A Technical Overview | p. 167 |
| Long PCR Methodology | p. 173 |
| Ultrasensitive and Quantitative PCR | |
| Qualitative and Quantitative PCR: A Technical Overview | p. 181 |
| Ultrasensitive PCR Detection of Tumor Cells in Myeloma | p. 185 |
| Ultrasensitive Quantitative PCR to Detect RNA Viruses | p. 197 |
| Quantitative PCR for cAMP RI Alpha mRNA: Use of Site-Directed Mutation and PCR Mimics | p. 205 |
| Quantitation of Multiple RNA Species | p. 211 |
| Transcriptome Analysis | |
| Differential Display: A Technical Overview | p. 217 |
| AU-Differential Display, Reproducibility of a Differential mRNA Display Targeted to AU Motifs | p. 225 |
| PCR Fluorescence Differential Display | p. 237 |
| Microarray Analysis Using RNA Arbitrarily Primed PCR | p. 245 |
| Oligonucleotide Arrays for Genotyping: Enzymatic Methods for Typing Single Nucleotide Polymorphisms and Short Tandem Repeats | p. 255 |
| Serial Analysis of Gene Expression | p. 271 |
| Mutations and Polymorphisms | |
| Mutation and Polymorphism Detection: A Technical Overview | p. 287 |
| Combining Multiplex and Touchdown PCR for Microsatellite Analysis | p. 295 |
| Detection of Microsatellite Instability and Loss of Heterozygosity Using DNA Extracted from Formalin-Fixed Paraffin-Embedded Tumor Material by Fluorescence-Based Multiplex Microsatellite PCR | p. 301 |
| Reaction of Shadow Band Synthesis During PCR Amplification of Repetitive Sequences from Modern and Ancient DNA | p. 309 |
| Degenerate Oligonucleotide-Primed PCR | p. 315 |
| Mutation Detection Using RT-PCR-RFLP | p. 319 |
| Multiplex Amplification Refractory Mutation System for the Detection of Prothrombotic Polymorphisms | p. 323 |
| PCR-SSCP Analysis of Polymorphism: A Simple and Sensitive Method for Detecting Differences Between Short Segments of DNA | p. 327 |
| PCR-Based Sequencing | |
| Sequencing: A Technical Overview | p. 337 |
| Preparation and Direct Automated Cycle Sequencing of PCR Products | p. 341 |
| Nonradioactive PCR Sequencing Using Digoxigenin | p. 347 |
| Direct Sequencing by Thermal Asymmetric PCR | p. 355 |
| Analysis of Nucleotide Sequence Variations by Solid-Phase Minisequencing | p. 361 |
| Direct Sequencing with Highly Degenerate and Inosine-Containing Primers | p. 367 |
| Determination of Unknown Genomic Sequences Without Cloning | p. 373 |
| Cloning PCR Products for Sequencing in M13 Vectors | p. 385 |
| DNA Rescue by the Vectorette Method | p. 393 |
| Technical Notes for Sequencing Difficult Templates | p. 401 |
| In Situ PCR and PRINS | |
| 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 | p. 405 |
| Cycling Primed In Situ Amplification | p. 425 |
| Direct and Indirect In Situ PCR | p. 433 |
| Reverse Transcriptase In Situ PCR: New Methods in Cellular Interrogation | p. 445 |
| Primed In Situ Nucleic Acid Labeling Combined with Immunocytochemistry to Simultaneously Localize DNA and Proteins in Cells and Chromosomes | p. 453 |
| Cloning and Mutagenesis | |
| Cloning and Mutagenesis: A Technical Overview | p. 467 |
| Using T4 DNA Polymerase to Generate Clonable PCR Products | p. 469 |
| A T-Linker Strategy for Modification and Directional Cloning of PCR Products | p. 475 |
| Cloning Gene Family Members Using PCR with Degenerate Oligonucleotide Primers | p. 485 |
| cDNA Libraries from a Low Amount of Cells | p. 499 |
| Creation of Chimeric Junctions, Deletions, and Insertions by PCR | p. 511 |
| Recombination and Site-Directed Mutagenesis Using Recombination PCR | p. 517 |
| Megaprimer PCR: Application in Mutagenesis and Gene Fusion | p. 525 |
| Index | p. 533 |
| Table of Contents provided by Publisher. All Rights Reserved. |
