Skip Navigation

This Article
Right arrow Print PDF (3732K)
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 Weinstock, P. H.
Right arrow Articles by Wetmur, J. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Weinstock, P. H.
Right arrow Articles by Wetmur, J. G.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1990, Vol. 18, No. 14 4207-4213
© 1990

CHEMISTRY

Branch capture reactions: effect of recipient structure

Peter H. Weinstock and James G. Wetmur*

Department of Microbiology, Box 1124, Mount Sinai School of Medicine New York, NY 10029, USA

*To whom correspondence should be addressed

Received March 20, 1990. Revised June 18, 1990. Accepted June 18, 1990.

Branch capture reactions (BCR) contain two DNA species: (i) a recipient restriction fragment terminating in an overhang and (ii) a displacer-linker duplex terminating in a displacer tail complementary to the overhang as well as contiguous nucleotides within the recipient duplex. Branched complexes containing both species are captured by ligation of the linker to the recipient overhang. Specificity depends upon branch migration and is increased by substitution of bromodeoxycytidine for deoxycytidine in the displacer. BCR rates and specificities were determined for recipient overhangs that were (i) 5‘ and 3’, (ii) 3 and 4 nucleotides long, and (iii) 0–100% G + C. Model systems permitted independent determination of G + C and branching effects on ligation rates and verification of rapid equilibrium between the branched complex and its component species. With all 4-base overhangs, recipient duplexes permitting extensive branch migration became saturated with displacer-linker duplexes. With increasing G + C, increasing ligation at competing sites led to decreased BCR specificity. BCR may be used to label a DNA fragment prior to electrophoresis, mark a fragment for affinity chromatography, or introduce a new overhang sequence compatible with a restriction endonuclease site in a cloning vector. A protocol was confirmed for mapping restriction sites in cloned DNA.


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
 Nucleic Acids ResHome page
B. I. Kankia
Optical absorption assay for strand-exchange reactions in unlabeled nucleic acids
Nucleic Acids Res., November 1, 2004; 32(19): e154 - e154.
[Abstract] [Full Text] [PDF]

Home page
 Genome Res.Home page
F Barany
The ligase chain reaction in a PCR world.
Genome Res., August 1, 1991; 1(1): 5 - 16.
[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.