home
Improved Cy3 and Cy5 DNA Labeling Efficiency

Linker-optimized Cy3- and Cy5-modified dUTPs
   



Please click here if this newsletter is not displayed properly! To contact us please DO NOT reply to this newsletter but send your inquiry to info@jenabioscience.com.


   
   

Labeling of DNA probes for in situ hybridization is most often performed by enzymatic incorporation of labeled dUTP using Taq DNA polymerase (= Polymerase Chain Reaction (PCR))[1-2].   More information...

We now offer linker-optimized Cy3- and Cy5-modified dUTPs (dUTP-XX-Cy3 and dUTP-XX-Cy5) that possess a longer linker resulting in higher labeling efficiencies at similar substitution rates (Fig. 1, Tab. 1)[3].


Figure 1: The longer the linker the higher the labeling efficiency at a similar dTTP substitution rate. PCR conditions: A 500 bp DNA fragment has been amplified from 10 ng genomic lambda DNA with Taq Polymerase. 10 µl probe were subsequently analyzed by agarose gel electrophoresis (1.8 % gel) without (A) or with DNA intercalator EvaGreen (B), M214: 100 bp DNA Ladder (Jena Bioscience), 0 % - 100 %: Increasing amount of dTTP substitution by Cy3-modified dUTP.


Table 1: Spectroscopic and substrate properties of Cy3- and Cy5-labeled dUTPs.

Nucleotide

Excmax
[nm] **

Emmax
[nm] **

εmax
[L x mmol-1 cm-1] **

Recommended dTTP
substitution by modified dUTP [%]
(PCR (Taq Pol)) *

dUTP-Cy3

550

570

150

30 - 50 %

dUTP-XX-Cy3

dUTP-Cy5

649

670

250

dUTP-XX-Cy5

20 - 40 %

* Please note: Optimal final concentration of the dye-labeled dUTP may vary depending on the application and assay conditions. For optimal product yields and optimal incorporation rates an individual optimization of the dye-labeled dUTP/ dTTP ratio is recommended.
** free acid


Jena Bioscience is a primary manufacturer offering


Selected References:
[1] Morrison et al. (2003) Labeling Fluorescence In Situ Hybridization Probes for Genomic Targets. Methods in Molecular Biology 204:21.
[2] Wiegant et al. (1997) Probe Labeling and Fluorescence In Situ Hybridization. In: Current Protocols in Cytometry 8.3.1, John Wiley & Sons Inc.
[3] Zhu et al. (1994) Directly labeled DNA probes using fluorescent nucleotides with different length linkers. Nucleic Acids Research 22:16.

   
   
   
   Responsible for content / Imprint
   
Jena Bioscience GmbH
Loebstedter Str. 71
07749 Jena
Germany

Phone: +49 – 3641 – 6285 000
Fax: +49 – 3641 – 6285 100
E-Mail: info@jenabioscience.com

Register Court: Amtsgericht Jena, HRB 207171
VAT No.: DE 195825742

        

Managing Directors:
Thomas Billert
Dr. Mathias Grün
   
    www.jenabioscience.com   
           
      © 2017 Jena Bioscience GmbH - Imprint