Protocols in Human Molecular Genetics

1. Front Matter

   Pages i-x

   PDF

2. The Polymerase Chain Reaction

   • Charles R. M. Bangham

   Pages 1-8

3. Direct DNA Sequencing of Complementary DNA Amplified by the Polymerase Chain Reaction

   • Richard A. Gibbs, Phi-Nga Nguyen, C. Thomas Caskey

   Pages 9-20

4. Direct Sequencing of PCR-Amplified DNA

   • Peter M. Green, Francesco Giannelli

   Pages 21-28

5. Rapid DNA Sequence Analysis Using Fluorescent Labels

   • Richard K. Wilson

   Pages 29-37

6. Detection of Mutations in DNA and RNA by Chemical Cleavage

   • R. G. H. Cotton

   Pages 39-49

7. Rapid Methods for Detection of Polymorphic Markers in Genomic DNA

   • David R. Bentley, Roland G. Roberts, Jane Montandon

   Pages 51-68

8. The Analysis of Point Mutations Using Synthetic Oligonucleotide Probes

   • Christine J. Farr

   Pages 69-76

9. Detection of Mutations by the Amplification Refractory Mutation System (ARMS)

   • John M. Old

   Pages 77-84

10. Automated Gene Detection Using the Oligonucleotide Ligation Assay

    • Ulf Landegren

    Pages 85-93

11. Detection of Point Mutations by Denaturing-Gradient Gel Electrophoresis

    • Stephen R. Dlouhy, Patricia Wheeler, James A. Trofatter, Peter J. Stambrook, Jay A. Tischfield

    Pages 95-110

12. The Detection and Mapping of Point Mutations by RNase A Cleavage

    • J. Ross Hawkins, Raymond Dalgleish

    Pages 111-121

13. Discontinuous Polyacrylamide Gel Electrophoresis of DNA Fragments

    • Bruce Budowle, Robert C. Allen

    Pages 123-132

14. Extraction and Enzymatic Amplification of DNA from Paraffin-Embedded Specimens

    • Colin S. Cooper, Michael R. Stratton

    Pages 133-140

15. The Use of the Polymerase Chain Reaction in the Mapping of Human Genes Using Somatic Cell Hybrids

    • Cathy Abbott, Sue Povey

    Pages 141-146

16. The Southern Blot

    • Michael R. Evans, Andrew L. Bertera, Dennis W. Harris

    Pages 147-157

17. The Detection of Specific DNA Sequences by Enhanced Chemiluminescence

    • Timothy C. Richardson, Ian Durrant

    Pages 159-168

18. Pulsed-Field Gel Electrophoresis

    • Johan T. den Dunnen, Gert-Jan B. van Ommen

    Pages 169-182

19. Cloning from Gels Following Pulse-Field Gel Electrophoresis

    • Peter J. Scambler, Michele Ramsay

    Pages 183-196

20. Yeast Artificial-Chromosome (YAC) Cloning Systems

    • Michele Ramsay, Carol Wicking

    Pages 197-221

Extraordinary advances have been made in the field of human molecular genetics during the past five years. The ability to amplify a specific region ofDNA a millionfold in a few hours using the polymerase chain reaction has led to the rapid identification of mutations in human disease and of DNA sequence polymorphisms on every human chro- some. DNA fragments of up to 1 megabase in length can now be resolved by pulsed-field gel electrophoresis to create long-range physical maps of important regions of the genome, and can be cloned in the form of yeast artificial chromosomes. The discovery of highly variable "minisatellite" DNA sequences has led to the development of DNA fingerprinting. The application of these techniques to the study of the human genome has culminated in major advances such as the cloning of the cystic fibrosis gene, the construction of genetic linkage maps of each human chro- some, the mapping of many genes responsible for human inherited d- orders, genetic fingerprinting of forensic specimens, and the detection of mutations involved in the development of human tumors. Although many of the new techniques in molecular genetics can be learned relatively easily, it is often difficult for a researcher to obtain all of the relevant information necessary for getting up a technique and applying it successfully. The information available in the research lite- ture often lacks the depth of description that the new user requires.

Cover[s] virtually all aspects of modern human molecular genetics....This is an excellent text and should be available to all laboratories using molecular biology as it offers precise instruction for the novice and a wide range of new techniques for the experienced researcher. Its popularity can be gauged by the intense interest shown in the book by both research scientists and students in our laboratory. -SGM Quarterly

• United Medical and Dental Schools of Guy’s, St. Thomas’s Hospitals, London, UK

  Christopher G. Mathew

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