Friday, October 18, 2013

Product Focus: 5-Azide-UTP: A novel approach for in vitro transcription-based RNA probe Labeling

The novel 5-UTP-Azidefrom Jena Bioscience provides a superior two-step approach for in vitro transcription-mediated RNA labeling based on Copper-free Click Chemistry.
It is available as a single nucleotide & as an optimized labeling kit and suitable for
  • preparation of non-radioactively labeled RNA probes (up to 1400 bp tested) (Fig. 1)
  • 3′-Azide-Labeling of T7 promotor-containing oligonucleotides[1,2]

Figure 1: An optimal balance between transcription efficiency and labeling density (2-3 dyes per 100 bp) is achieved with 25 – 50 % 5-Azide-UTP. Azide-functionalized RNA was prepared by T7 RNA pol-mediated in vitro transcription with increasing 5-Azide-UTP concentrations (0 – 100 %, 200 ng of a 1423 bp DNA template, 1 h, 37°C), purified and subsequently labeled with DBCO-Sulfo-Cy3 via Copper-free Click Chemistry. Transcripts were analyzed by gel electrophoresis: A) Cy3-labeled RNA without EtBr (Cy3-signal, labeling density) and B) RNA prior to labeling in the presence of EtBr (EtBr-signal, transcription efficiency). L= ladder marker.


Where can I find more information about AAT Bioquest?

Visit the manufacturer page at www.stratech.co.uk/aatbioquest, email info@stratech.co.uk or call +44 (0) 1638 782600.
Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.
Post a Comment