Skip Navigation

This Article
Right arrow Print PDF (5392K)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Spanos, A.
Right arrow Articles by Banks, G. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Spanos, A.
Right arrow Articles by Banks, G. R.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1981, Vol. 9, No. 8 1825-1839
© 1981

ENZYMOLOGY

Detection of the catalytic activities of DNA polymerases and their associated exonucleases following SDS-polyacrylamide gel electrophoresis

Ad Spanos, Steven G. Sedgwick, Geoffrey T. Yarranton, Ulrich Hübscher and Geoffrey R. Banks

Received February 16, 1981. A method is described to detect DNA polymerases and nucleases in homogeneous or crude enzyme preparations after electrophoresis in SDS-polyacrylamide gels(2) containing the appropriate template or substrate. DNA polymerases are electrophoresed in a gel containing gapped calf thymus DNA and after a renaturation treatment, the gel is incubated in a reaction mixture in which one deoxyribonucleoside triphosphate is [{propto}-32P]-labelled. Incorporation of radioactivity into DNA is detected at the vicinity of the polymerase band by autoradiography. An associated nuclease activity can be measured after electrophoresis in a gel containing 32P -labelled gapped DNA, when nucleolytic digestion is seen as a clear band in the resulting autoradiogram. The gels can subsequently be stained with Coomassie blue to establish identical molecular weights of polymerase, nuclease and protein bands. Applications of this technique are discussed.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
K. Ramadan, I. V. Shevelev, G. Maga, and U. Hubscher
DNA Polymerase lambda from Calf Thymus Preferentially Replicates Damaged DNA
J. Biol. Chem., May 17, 2002; 277(21): 18454 - 18458.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S. L. Washington, M. S. Yoon, A. M. Chagovetz, S.-X. Li, C. A. Clairmont, B. D. Preston, K. A. Eckert, and J. B. Sweasy
A genetic system to identify DNA polymerase beta  mutator mutants
PNAS, February 18, 1997; 94(4): 1321 - 1326.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
A. W. Plug, C. A. Clairmont, E. Sapi, T. Ashley, and J. B. Sweasy
Evidence for a role for DNA polymerase beta  in mammalian meiosis
PNAS, February 18, 1997; 94(4): 1327 - 1331.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. Shinkai, P. H. Patel, and L. A. Loeb
The Conserved Active Site Motif A of Escherichia coli DNA Polymerase I Is Highly Mutable
J. Biol. Chem., May 25, 2001; 276(22): 18836 - 18842.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.