UV cross-linking of unmodified DNA on glass surfaces.
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.
Schüler
Thomas
T
Jenaer Biochip Initiative, Institute of Physical Chemistry, Friedrich Schiller University, Helmholtzweg 4, 07743 Jena, Germany.
Nykytenko
Alla
A
Csaki
Andrea
A
Möller
Robert
R
Fritzsche
Wolfgang
W
Popp
Jürgen
J
D004247:DNA / Q000737:chemistry* / Q000528:radiation effects*; D005898:Glass / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis; D012680:Sensitivity and Specificity; D012821:Silanes / Q000737:chemistry*; D013499:Surface Properties; D014466:Ultraviolet Rays
D012821:Silanes; D004247:DNA