UID:
edocfu_9960073333202883
Format:
1 online resource (397 p.)
ISBN:
1-281-75638-5
,
9786611756383
,
0-08-085879-1
Series Statement:
Laboratory techniques in biochemistry and molecular biology ; 10
Content:
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.
Note:
Description based upon print version of record.
,
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
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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
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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
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5.3. Separating the two labelled ends of complementary strands5.4. 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
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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
,
English
Additional Edition:
ISBN 0-444-80497-8
Language:
English
