DNA Sequencing Protocols

1. Front Matter

   Pages i-xii

   PDF

2. DNA Sequencing

   • Hugh G. Griffin, Annette M. Griffin

   Pages 1-8

3. M13 Cloning Vehicles

   • Joachim Messing

   Pages 9-22

4. Cloning into M13

   • Qingzhong Yu

   Pages 23-30

5. Transfection of E. coli with M13 DNA

   • Fiona M. Tomely

   Pages 31-36

6. M13 Phage Growth and Single-Strand DNA Preparation

   • Fiona M. Tomley

   Pages 37-40

7. M13 Phage Growth and DNA Purification Using 96 Well Microtiter Trays

   • Alan T. Bankier

   Pages 41-46

8. Generation of Random Fragments by Sonication

   • Alan T. Bankier

   Pages 47-50

9. Generation of a Nested Set of Deletions Using Exonuclease III

   • George Murphy

   Pages 51-60

10. Sequential Deletions of Single-Stranded DNA

    • George Murphy

    Pages 61-68

11. Subcloning for DNA Sequencing

    • Gary M. Studnicka, Shau-Ping Lei, Hun-Chi Lin, Gary Wilcox

    Pages 69-74

12. Sequencing Using Custom Designed Oligonucleotides

    • Ulrike Gerischer, Peter Dürre

    Pages 75-82

13. Dideoxy Sequencing Reactions Using Klenow Fragment DNA Polymerase 1

    • Alan T. Murphy

    Pages 83-90

14. Dideoxy Sequencing Reactions Using T7 Polymerase

    • Peter Dürre, Ulrike Geriscker

    Pages 91-102

15. Dideoxy Sequencing Reactions Using Sequenase Version 2.0

    • Hugh G. Griffin, Annette M. Griffin

    Pages 103-108

16. Dideoxy Sequencing Reactions Using Taq Polymerase

    • Fabrizio Arigoni, P. Alexandre Kaminski

    Pages 109-114

17. Pouring Linear and Buffer-Gradient Sequencing Gels

    • Paul Littlebury

    Pages 115-120

18. Electrophoresis of Sequence Reaction Samples

    • Alan T. Bankier

    Pages 121-130

19. Plasmid Sequencing

    • Hugh G. Griffin, Annette M. Griffin

    Pages 131-136

20. Plasmid Sequencing

    • George Murphy

    Pages 137-140

The purpose of DNA Sequencing Protocols is to provide detailed practical procedures for the widest range of DNA sequencing meth­ ods, and we believe that all the vanguard techniques now being applied in this fast-evolving field are comprehensively covered. Sequencing technology has advanced at a phenomenal rate since the original methods were first described in the late 1970s and there is now a huge variety of strategies and methods that can be employed to determine the sequence of any DNA of interest. More recently, a large number of new and innovative sequencing techniques have been developed, including the use of such novel polymerases as Tag poly­ merase and Sequenase, the harnessing of PCR technology for linear amplification (cycle) sequencing, and the advent of automated DNA sequencers. DNA sequencing is surely one of the most important techniques in the molecular biology laboratory. Sequence analysis is providing an increasingly useful approach to the characterization of biological systems, and major multinational projects are already underway to map and sequence the entire genome of organisms, such as Escherichia coli, Saccharomyces cerevisiae, Caenorhabditis elegans, and Homo sapiens. Most scientists recognize the importance of DNA sequence data and perceive DNA sequencing as a valuable and indispensable aspect of their work. Recent technological advances, especially in the area of automated sequencing, have removed much of the drudg­ ery that was formerly associated with the technique, whereas innova­ tive computer software has greatly simplified the analysis and manipulation of sequence data.

...38 chapters, all of the greatest interest...an indispensable book of the highest practical use.-Cellular and Molecular Biology

• Institute of Food Research, Norwich Research Park, Norwich, England

  Hugh G. Griffin, Annette M. Griffin

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