An inexpensive and simple method for thermally stable immobilization of DNA on an unmodified glass surface: UV linking of poly(T)10-poly(C)10-tagged DNA probes.
Microarrays printed on glass slides are often constructed by covalently linking modified oligonucleotide probes to a derivatized surface at considerable expense. In this article, we demonstrate that 14-base oligonucleotides with a poly(T)10 - poly(C)10 tail (TC tag), but otherwise unmodified, can be linked by UV light irradiation onto a plain, unmodified glass surface. Probes immobilized onto unmodified glass microscope slides performed similarly to probes bound to commercial amino-silane-coated slides and had comparable detection limits. The TC-tagged probes linked to unmodified glass did not show any significant decrease in hybridization performance after a 20 min incubation in water at 100 degrees C prior to rehybridization, indicating a covalent bond between the TC tag and unmodified glass. The probes were used in thermal minisequencing cycling reactions. Furthermore, the TC tag improved the hybridization performance of the immobilized probes on the amino-silane surface, indicating a general benefit of adding a TC tag to DNA probes. In conclusion, our results show that using TC-tagged DNA probes immobilized on an unmodified glass surface is a robust, heat-stable, very simple, and inexpensive method for manufacturing DNA microarrays.
Gudnason
Haukur
H
DTU Nanotech, Department of Micro- and Nanotechnology, Technical University of Denmark, Lyngby.
Dufva
Martin
M
Duong Bang
Dang
D
Wolff
Anders
A
D004247:DNA / Q000235:genetics*; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000235:genetics* / Q000528:radiation effects; D005898:Glass / Q000737:chemistry*; D006358:Hot Temperature; D006801:Humans; D007103:Immobilization; D009693:Nucleic Acid Hybridization / Q000379:methods; D011066:Poly C / Q000235:genetics* / Q000528:radiation effects; D011071:Poly T / Q000235:genetics* / Q000528:radiation effects; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D013499:Surface Properties; D014466:Ultraviolet Rays
D017931:DNA Primers; D015342:DNA Probes; D011071:Poly T; D011066:Poly C; D004247:DNA