SpringerProtocols: Abstract: Dideoxy Sequencing Reactions Using Klenow Fragment DNA Polymerase 1

Contents of this article

Browse by Subject

Upload a Protocol

Upload your own protocols for personal use.

Protocol Alert

Receive e-mail notifications about new content on SpringerProtocols.

Video Protocols

View our video protocols.

Call for video protocols.

Comments

Read comments by other users and add your own.

Favorites

Save your favorite protocols to your My Protocols area.

RSS

RSS Feeds

Inside SpringerProtocols

Useful Tools

Dideoxy Sequencing Reactions Using Klenow Fragment DNA Polymerase 1

By: Alan T. Murphy²

Abstract

Full Text

Download PDF (485K)

Historically, DNA sequencing by primed synthesis and chain terminators has been performed using the Klenow fragment of DNA polymerase 1(1. This enzyme was derived from E. coli. DNA Pol I which, in addition to polymerase activity, has both 3′–5′ and 5′–3′ exonuclease activities.

Affiliation(s): (2) MRC Lab of Molecular Biology, Cambridge, England

Book Title: DNA Sequencing Protocols

Series: Methods in Molecular Biology | Volume: 23 | Pub. Date: Aug-23-1993 | Page Range: 83-90 | DOI: 10.1385/0-89603-248-5:83

Subject: Genetics/Genomics

References

Download References

1.	Sanger, F., Nicklen, S, and Coulson, A. R. (1977) DNA sequencing with chainterminating inhibitors Proc Nat1 Acad. Sci USA 74, 5463–54

2.	Bankier, A. T. and Barrell, B. G(1983) Shotgun DNA sequencing, rn Techniques in Life Sciences, Nucleic Acid Biochemistry vol. B5, (Flavell, R. A., ed.), Elsevier Scientific Publishers, Ireland, pp. 1–34.

3.	Bankier, A T, Weston, K. M, and Barrell, B G (1987) Random cloning and sequencing by the M13/dideoxynucleotide chain termination method, in Methods in Enzymology vol. 155, (Wu, R., ed.), Academic, London, pp. 51–93

4.	Bankrer, A. T. and Barrell, B. G. (1989) Sequencing single-stranded DNA using the chain termination method, in Nuclerc Acids Sequencrng. A Practical Approach (Howe, C. J. and Ward, E. S., eds.), IRL Press, Oxford, pp. 37–78

5.	Messing, J. and Bankier, A T (1989) The use of single-stranded DNA phage in DNA sequencing, in Nucleic Acids Sequencing. A Practical Approach (Howe, C. J. and Ward, E. S., eds.), IRL Press, Oxford, pp 1–36

6.	Biggin, M. D, Gibson, T. J., and Hong, G. F. (1983) Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination Proc. Natl. Acad. Sci. USA 80, 3963–3965.

7.	Zagursky, R J., Conway, P S, and Kashdan, M A. (1991) Use of 33P for Sanger DNA Sequencing Biotechniques 1, 37–38

Comments (Loading...)

Post comment/View all comments