Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (188K) Freely available
Right arrow Supplementary Material
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 Marras, S. A. E.
Right arrow Articles by Tyagi, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Marras, S. A. E.
Right arrow Articles by Tyagi, S.
Related Collections
Right arrow Monitoring gene expression
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Published online 20 May 2004

Nucleic Acids Research, 2004, Vol. 32, No. 9 e72
© 2004 Oxford University Press

Real-time measurement of in vitro transcription

Salvatore A. E. Marras*, Benjamin Gold, Fred Russell Kramer, Issar Smith and Sanjay Tyagi

Public Health Research Institute, 225 Warren Street, Newark, NJ 07103, USA

*To whom correspondence should be addressed. Tel: +1 973 854 3373; Fax: +1 973 854 3374; Email: marras{at}phri.org
Present address:
Benjamin Gold, Department of Microbiology and Immunology, Weill Medical College of Cornell University, New York, NY 10021, USA
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors

Received March 23, 2004; Revised and Accepted April 16, 2004

We have developed a simple method to measure RNA synthesis in real time. In this technique, transcription reactions are performed in the presence of molecular beacons that possess a 2'-O-methylribonucleotide backbone. These probes become fluorescent as they hybridize to nascent RNA during the course of synthesis. We found that molecular beacons synthesized from natural deoxyribonucleotides were not suitable, because they are copied by RNA polymerases, generating complementary product strands that bind to the molecular beacons, causing a conformational change that results in unwanted fluorescence. However, when the molecular beacons are synthesized from 2'-O-methylribonucleotides, they are not copied and fluorescence is strictly dependent upon transcription of the added template. Utilizing these modified molecular beacons, quantitative comparisons were made of the activity of a variety of RNA polymerases and the effect of an inhibitor of transcription was determined.


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
K. Hosoda, T. Matsuura, H. Kita, N. Ichihashi, K. Tsukada, I. Urabe, and T. Yomo
A novel sequence-specific RNA quantification method using nicking endonuclease, dual-labeled fluorescent DNA probe, and conformation-interchangeable oligo-DNA
RNA, March 1, 2008; 14(3): 584 - 592.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
Y. Kim, C. J. Yang, and W. Tan
Superior structure stability and selectivity of hairpin nucleic acid probes with an L-DNA stem
Nucleic Acids Res., December 18, 2007; 35(21): 7279 - 7287.
[Abstract] [Full Text] [PDF]

Home page
 DevelopmentHome page
M. Kloc, K. Wilk, D. Vargas, Y. Shirato, S. Bilinski, and L. D. Etkin
Potential structural role of non-coding and coding RNAs in the organization of the cytoskeleton at the vegetal cortex of Xenopus oocytes
Development, August 1, 2005; 132(15): 3445 - 3457.
[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.