Biology Reference
In-Depth Information
CHAPTER 8
DNA Amplification by the
Polymerase Chain Reaction:
Molecular Biology Made Accessible
Chapter Outline
8.1 Overview 308
8.2 Introduction 309
8.3 The Basic Polymerase Chain Reaction (PCR)
312
8.3.1
The First Few Cycles are Critical
312
8.3.2 PCR Power 313
8.3.3
Standard PCR Protocols
313
8.3.4 DNA Polymerases 313
8.3.5
Other Thermostable DNA Polymerases
315
8.3.6
Primers are Primary
315
8.3.7
Storing Insects for the PCR
318
8.3.8
Preparing DNA Samples
319
8.3.9 PCR Automation 324
8.3.10 Specificity of the PCR 324
8.3.11 Detecting Primer Artifacts 326
8.3.12 How Many Cycles Does a PCR Need?
326
8.3.13 Reducing the Evils of Contamination
327
8.4 Some Modifications of the PCR
329
8.4.1
AFLP for DNA Fingerprinting
329
8.4.2 Anchored PCR 333
8.4.3 Arbitrary Primers 333
8.4.4 Asymmetric PCR 334
8.4.5 Degenerate Primers 334
8.4.6 Hot-Start PCR 335
8.4.7 Inverse PCR 335
8.4.8
Long PCR or High-Fidelity PCR
336
8.4.9 Multiplex PCR 339
8.4.10 Nested PCR 341
8.4.11 PCR-RFLP 341
8.4.12 Quantitative PCR
342
