Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (392K) Freely available
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 arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Allers, T.
Right arrow Articles by Lichten, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Allers, T.
Right arrow Articles by Lichten, M.
Related Collections
Right arrow Recombination
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2000, Vol. 28, No. 2 E6-e6
© 2000 Oxford University Press

A method for preparing genomic DNA that restrains branch migration of Holliday junctions

Thorsten Allers and Michael Lichten*

Laboratory of Biochemistry, Division of Basic Sciences, Building 37, Room 4C03, National Cancer Institute, 37 Convent Drive, Bethesda, MD 20892-4255, USA

The Holliday junction is a central intermediate in genetic recombination. This four-stranded DNA structure is capable of spontaneous branch migration, and is lost during standard DNA extraction protocols. In order to isolate and characterize recombination intermediates that contain Holliday junctions, we have developed a rapid protocol that restrains branch migration of four-way DNA junctions. The cationic detergent hex-adecyltrimethylammonium bromide is used to lyse cells and precipitate DNA. Manipulations are performed in the presence of the cations hexamine cobalt(III) or magnesium, which stabilize Holliday junctions in a stacked-X configuration that branch migrates very slowly. This protocol was evaluated using a sensitive assay for spontaneous branch migration, and was shown to preserve both artificial Holliday junctions and meiotic recombination intermediates containing four-way junctions.

* To whom correspondence should be addressed. Tel: +1 301 496 3393; Fax: +1 301 402 7437; Email: lichten@helix.nih.gov


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
 Mol. Biol. CellHome page
H. W. Mankouri, H.-P. Ngo, and I. D. Hickson
Shu Proteins Promote the Formation of Homologous Recombination Intermediates That Are Processed by Sgs1-Rmi1-Top3
Mol. Biol. Cell, October 1, 2007; 18(10): 4062 - 4073.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Fierro-Fernandez, P. Hernandez, D. B. Krimer, and J. B. Schvartzman
Replication Fork Reversal Occurs Spontaneously after Digestion but Is Constrained in Supercoiled Domains
J. Biol. Chem., June 22, 2007; 282(25): 18190 - 18196.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
M. Fierro-Fernandez, P. Hernandez, D. B. Krimer, A. Stasiak, and J. B. Schvartzman
From the Cover: Topological locking restrains replication fork reversal
PNAS, January 30, 2007; 104(5): 1500 - 1505.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Biol. CellHome page
H. W. Mankouri and I. D. Hickson
Top3 Processes Recombination Intermediates and Modulates Checkpoint Activity after DNA Damage
Mol. Biol. Cell, October 1, 2006; 17(10): 4473 - 4483.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
F. Prado and A. Aguilera
Partial Depletion of Histone H4 Increases Homologous Recombination-Mediated Genetic Instability
Mol. Cell. Biol., February 15, 2005; 25(4): 1526 - 1536.
[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.