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This handbook, designed for geneticists and molecular biologists, offers a variety of methods for DNA sequencing suitable for human genome sequencing and mutation detection in human disease. Step-by-step instructions and notes addressing potential problems are included for methods such as automated DNA sequencing, capillary DNA sequencers, heterozygote mutation detection, web-based sequencing databases, and genome sequencing sites. Also included is a review of shotgun sequencing in the human genome project. Spiral binding.

Book Info

Belfast City Hospital Trust, Northern Ireland. Provides detailed practical procedures for a range of the most commonly applied DNA sequencing methods. Strives to pinpoint those areas within each of the techniques that cause the most problems. Previous edition: c1993. Plastic-comb binding, hardcover also available.

Preface v
Contributors ix
1 Introduction to DNA Sequencing 1
2 The Universal Primers and the Shotgun DNA Sequencing Method 13
3 M13 Sequencing 33
4 Primer Design and Primer-Directed Sequencing 39
5 Direct Sequencing of DNA Produced in a Polymerase Chain Reaction 53
6 Solid Phase Fluorescent Sequencing of the CFTR Gene 63
7 Shotgun DNA Sequencing 89
8 Cycle Sequencing 101
9 Cycle Sequencing of Polymerase Chain Reaction-Amplified Genomic DNA with Dye-Labeled Universal Primers 113
10 Automated Fluorescent DNA Sequencing on the ABI PRISM 377 119
11 Automated Fluorescent DNA Sequencing on the ABI PRISM 310 Genetic Analyzer 153
12 Fluorescent Sequencing Protocols for the ALF 171
13 Fluorescent Sequencing for Heterozygote Mutation Detection 193
14 Sequence Databases and the Internet 215
15 DNA Sequencing by Capillary Array Electrophoresis 225
Index 241