DNA Sequencing
Published on 1. April 2000
383 pages
978-0-08-085879-1 (ISBN)
Description
This volume provides a comprehensive description of the principles and methods used in DNA sequencing. Following a detailed introduction the chapters are: DNA sequencing; Chain terminator sequencing; Primed synthesis methods applied to DNA fragments cloned into phage M13; DNA sequencing by the Maxam-Gilbert chemical procedure; Computer methods for DNA sequencers; Appendices including contractions and special terms, cloning vectors, commercially available restriction endonucleases, and autoradiography.
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J. Hindley
DNA Sequencing: Volume 10
Book
04/2000
Elsevier
€86.65
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Content
- Front Cover
- DNA Sequencing
- Copyright Page
- Contents
- Acknowledgements
- Chapter 1. Introduction
- 1.1. Preliminary remarks
- 1.2. Organisation of the book
- 1.3. General background to DNA sequencing
- Chapter 2. DNA Sequencing
- 2.1. DNA sequencing by primed synthesis methods
- 2.2. Detailed protocol for the 'plus' and 'minus' primed synthesis method
- 2.3. Additional methods useful in conjunction with plus and minus method
- 2.4. Discussion and summary
- 2.5. Sequence analysis of short DNA fragments
- Chapter 3. Chain terminator sequencing
- 3.1. DNA sequencing with chain terminating inhibitors (Sanger et al., 1977)
- 3.2. Sequencing by partial ribosubstitution
- 3.3. Direct enzymatic method for sequencing double-stranded restriction fragments using dideoxynucleoside triphosphates (Maat and Smith. 1978)
- 3.4. The use of exonuclease Ill for preparing single-stranded DNA for use as a template in the chain terminator sequencing method (Smith, 1979, 1980)
- Chapter 4. Primed synthesis methods applied to DNA fragments cloned into phage M13
- 4.1. General introduction
- 4.2. Rapid random sequencing by shotgun cloning into single stranded phage vectors (Sanger et al., 1980)
- 4.3. Detailed experimental procedures
- Experimental procedures
- 4.4. Dideoxy-nucleotide sequencing procedures
- 4.5. Results
- 4.6. Discussion
- 4.7. Hints for reading sequencing gels
- 4.8. Sequencing long single-stranded DNAs cloned in bacteriophage M13 using internal primers
- 4.9. Sequencing cDNA from reverse transcripts of RNA
- Key references
- Chapter 5. DNA sequencing by the Maxam-Gilbert chemical procedure
- 5.1. Introduction
- 5.2. End-labelling DNA segments
- 5.3. Separating the two labelled ends of complementary strands
- 5.4. Strand separation
- 5.5. Elution of DNA from polyacrylamide gels
- 5.6. Base-specific chemical cleavage reactions
- 5.7. Gel electrophoresis
- 5.8. Reading the gel
- 5.9. Experimental procedures
- 5.10. Sequencing reactions (Maxam and Gilbert, 1980)
- 5.11. Trouble-shooting guide (Maxam and Gilbert, 1980)
- 5.12. Results
- 5.13. Strategies for sequencing double-stranded DNA
- Update
- Update to chapter 4
- Update to chapter 5
- Appendices
- Appendix 1. List of contractions and special terms
- Appendix 2. Cloning vectors
- Appendix 3. Commercially obtainable restriction endonucleases (1981)
- Appendix 4. Autoradiography
- Appendix 5. Equipment
- Appendix 6
- Appendix 7. Nucleotide sequence of M13mp7
- 7238 residues
- Appendix 8. Restriction map of M13mp8 (7229 base pairs)
- Chapter 6. Computer methods for DNA sequencers
- 1. Introduction
- 2. Data storage and retrieval: the DB system
- 3. Analysis: restriction sites, hairpin loops, translation, homologies
- 4. Choosing hardware and software
- 5. References
- Appendix to chapter 6
- Getting started with the DB system
- Running the programs
- References
- Subject index
