UV cross-linking of unmodified DNA on glass surfaces |
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Authors: | Thomas Schüler Alla Nykytenko Andrea Csaki Robert Möller Wolfgang Fritzsche Jürgen Popp |
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Institution: | 1.Jenaer Biochip Initiative, Institute of Physical Chemistry,Friedrich Schiller University,Jena,Germany;2.Institute of Photonic Technology,Jena,Germany |
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Abstract: | The performance of DNA microarrays strongly depends on their surface properties. Furthermore, the immobilization method of
the capture molecules is of importance for the efficiency of the microarray in terms of sensitivity and specificity. This
work describes the immobilization of single-stranded capture oligonucleotides by UV cross-linking on silanated (amino and
epoxy) glass surfaces. Thereby we used amino (NH2) and poly thymine/poly cytosine modifications of the capture sequences as well as unmodified capture molecules. The results
were compared to UV cross-linking of the same DNA oligonucleotides on unmodified glass surfaces. Immobilization and hybridization
efficiency was demonstrated by fluorescence and enzyme-induced deposition of silver nanoparticles. We found out that single-stranded
DNA molecules do not require a special modification to immobilize them by UV cross-linking on epoxy- or amino-modified glass
surfaces. However, higher binding rates can be achieved when using amino-modified oligonucleotides on an epoxy surface. The
limit of detection for the used settings was 5 pM. |
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