Characterization of a polymeric adsorbed coating for DNA microarray glass slides.
A new method was developed to covalently attach target molecules onto the surface of glass substrates such as microwell plates, beads, tubes, and microscope slides, for hybridization assays with fluorescent targets. The innovative concept introduced by this work is to physically adsorb onto underivatized glass surfaces a functional copolymer, able to graft amino-modified DNA molecules. The polymer, obtained by radical copolymerization of N,N-dimethylacrylamide, N-acryloyloxysuccinimide, and 3-(trimethoxysilyl)propyl methacrylate, copoly(DMA-NAS-MAPS), self-adsorbs onto the glass surface very quickly, typically in 5-30 min. The film, formed on the surface, bears active esters, which react with amino-modified DNA targets. The surface layer is stable in an aqueous buffer containing various additives (SDS, urea, salt), even at boiling temperature. It should be emphasized that the coating is formed by the immersion of glass slides in a diluted aqueous solution of the polymer. Therefore, the procedure is fast, inexpensive, robust, and reliable, and it does not require time-consuming glass pretreatments. Slides, coated with copoly(DMA-NAS-MAPS), were profitably used as substrates for the preparation of low-density DNA microarrays. The density and the thickness of the films were evaluated by X-ray reflectivity measurements whereas the extent of reaction of functional groups with DNA molecules was determined by a functional test. The experiments indicate that half of the active groups present on the surface reacts with oligonucleotide probes.
Pirri
Giovanna
G
Istituto di Chimica del Riconoscimento Molecolare, CNR, Milan, Italy.
Damin
Francesco
F
Chiari
Marcella
M
Bontempi
Elza
E
Depero
Laura E
LE
D000178:Acrylamides / Q000737:chemistry; D000327:Adsorption; D005898:Glass; D015394:Molecular Structure; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D009841:Oligonucleotides / Q000032:analysis; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry
D000178:Acrylamides; D009841:Oligonucleotides; D011108:Polymers; D012997:Solvents; C099046:N,N-dimethylacrylamide