Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (112K) 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 ISI Web of Science
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 Search for citing articles in:
ISI Web of Science (33)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Nilsson, M.
Right arrow Articles by Landegren, U.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Nilsson, M.
Right arrow Articles by Landegren, U.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2001, Vol. 29, No. 2 578-581
© 2001 Oxford University Press

RNA-templated DNA ligation for transcript analysis

Mats Nilsson*, Dan-Oscar Antson1, Gisela Barbany1 and Ulf Landegren1

Department of Molecular Cell Biology, Leiden University Medical Center, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands and 1Rudbeck Laboratory, Department of Genetics and Pathology, SE-75185 Uppsala, Sweden

Ligase-mediated gene detection has proven valuable for detection and precise distinction of DNA sequence variants. We have recently shown that T4 DNA ligase can also be used to distinguish single nucleotide variants of RNA sequences. Here we describe parameters that influence RNA-templated DNA ligation by T4 DNA ligase. The reaction proceeds much more slowly, requiring more enzyme, compared to ligation of the same oligonucleotides hybridized to the corresponding DNA sequence. The reaction is inhibited at high concentrations of ATP and NaCl and both magnesium and manganese ions can support the reaction. We define reaction conditions where 80% of RNA target molecules can template a diagnostic ligation reaction. Ligase-mediated RNA detection should provide a useful mechanism for sensitive and accurate detection and distinction of RNA sequence variants.

* To whom correspondence should be addressed. Tel: +31 71 5276194; Fax: +31 71 5276180; Email: m.nilsson{at}lumc.nl


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
 RNAHome page
A. Lagunavicius, E. Merkiene, Z. Kiveryte, A. Savaneviciute, V. Zimbaite-Ruskuliene, T. Radzvilavicius, and A. Janulaitis
Novel application of Phi29 DNA polymerase: RNA detection and analysis in vitro and in situ by target RNA-primed RCA
RNA, May 1, 2009; 15(5): 765 - 771.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
J. J. Li, Y. Chu, B. Y.-H. Lee, and X. S. Xie
Enzymatic signal amplification of molecular beacons for sensitive DNA detection
Nucleic Acids Res., April 1, 2008; 36(6): e36 - e36.
[Abstract] [Full Text] [PDF]

Home page
 RNAHome page
A. Lagunavicius, Z. Kiveryte, V. Zimbaite-Ruskuliene, T. Radzvilavicius, and A. Janulaitis
Duality of polynucleotide substrates for Phi29 DNA polymerase: 3'->5' RNase activity of the enzyme
RNA, March 1, 2008; 14(3): 503 - 513.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
J. M. Goodenbour and T. Pan
Diversity of tRNA genes in eukaryotes
Nucleic Acids Res., December 4, 2006; 34(21): 6137 - 6146.
[Abstract] [Full Text] [PDF]

Home page
 RNAHome page
Y. Wang and S. K. Silverman
Efficient RNA 5'-adenylation by T4 DNA ligase to facilitate practical applications
RNA, June 1, 2006; 12(6): 1142 - 1146.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Microbiol.Home page
B. Wang, S. J. Potter, Y. Lin, A. L. Cunningham, D. E. Dwyer, Y. Su, X. Ma, Y. Hou, and N. K. Saksena
Rapid and Sensitive Detection of Severe Acute Respiratory Syndrome Coronavirus by Rolling Circle Amplification
J. Clin. Microbiol., May 1, 2005; 43(5): 2339 - 2344.
[Abstract] [Full Text] [PDF]

Home page
 Genome ResHome page
J.-B. Fan, J. M. Yeakley, M. Bibikova, E. Chudin, E. Wickham, J. Chen, D. Doucet, P. Rigault, B. Zhang, R. Shen, et al.
A Versatile Assay for High-Throughput Gene Expression Profiling on Universal Array Matrices
Genome Res., May 1, 2004; 14(5): 878 - 885.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. Mendel-Hartvig, A. Kumar, and U. Landegren
Ligase-mediated construction of branched DNA strands: a novel DNA joining activity catalyzed by T4 DNA ligase
Nucleic Acids Res., January 2, 2004; 32(1): e2 - e2.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. Nilsson, M. Gullberg, F. Dahl, K. Szuhai, and A. K. Raap
Real-time monitoring of rolling-circle amplification using a modified molecular beacon design
Nucleic Acids Res., July 15, 2002; 30(14): e66 - e66.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
A. T. Christian, M. S. Pattee, C. M. Attix, B. E. Reed, K. J. Sorensen, and J. D. Tucker
Detection of DNA point mutations and mRNA expression levels by rolling circle amplification in individual cells
PNAS, November 20, 2001; (2001) 251383598.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
A. T. Christian, M. S. Pattee, C. M. Attix, B. E. Reed, K. J. Sorensen, and J. D. Tucker
Detection of DNA point mutations and mRNA expression levels by rolling circle amplification in individual cells
PNAS, December 4, 2001; 98(25): 14238 - 14243.
[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.