Skip Navigation

Nucleic Acids Research 2004 32(18):e140; doi:10.1093/nar/gnh138
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (282K) 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 Dekker, N. H.
Right arrow Articles by Dekker, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Dekker, N. H.
Right arrow Articles by Dekker, C.
Related Collections
Right arrow RNA characterisation and manipulation
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Published online 8 October 2004

Nucleic Acids Research, Vol. 32 No. 18 © Oxford University Press 2004; all rights reserved

Joining of long double-stranded RNA molecules through controlled overhangs

N. H. Dekker*, J. A. Abels, P. T. M. Veenhuizen, M. M. Bruinink and C. Dekker

Kavli Institute of Nanoscience, Faculty of Applied Sciences, Delft University of Technology, Lorentzweg 1, 2628 CJ Delft, The Netherlands

* To whom correspondence should be addressed. Tel: +31 15 2783219; Fax: +31 15 2781202; Email: Nynke.Dekker{at}mb.tn.tudelft.nl

Received August 10, 2004; Revised and Accepted September 22, 2004

We describe two methods for creating long (>1 kb) dsRNA molecules with specific, user-controlled overhangs for efficient hybridization and ligation. The two methods create double-stranded RNA (dsRNA) molecules with 5' overhangs or with 3' overhangs using T7 RNA polymerase (T7 RNAP) in transcription reactions of carefully designed PCR products. Primers utilized in the PCR reactions provide the template for the desired dsRNA overhangs. These methods provide complete control of the length and the sequence of the overhangs. This supplies a tool which is particularly lacking in dsRNA biochemistry given the absence of restriction endonucleases active on these substrates.


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
O. R. Faridani, G. M. McInerney, K. Gradin, and L. Good
Specific ligation to double-stranded RNA for analysis of cellular RNA::RNA interactions
Nucleic Acids Res., September 1, 2008; 36(16): e99 - e99.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
D. A. Koster, C. H. Wiggins, and N. H. Dekker
Multiple events on single molecules: Unbiased estimation in single-molecule biophysics
PNAS, February 7, 2006; 103(6): 1750 - 1755.
[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.