Functionalization of poly(methyl methacrylate) (PMMA) as a substrate for DNA microarrays.
A chemical procedure was developed to functionalize poly(methyl methacrylate) (PMMA) substrates. PMMA is reacted with hexamethylene diamine to yield an aminated surface for immobilizing DNA in microarrays. The density of primary NH2 groups was 0.29 nmol/cm2. The availability of these primary amines was confirmed by the immobilization of DNA probes and hybridization with a complementary DNA strand. The hybridization signal and the hybridization efficiency of the chemically aminated PMMA slides were comparable to the hybridization signal and the hybridization efficiency obtained from differently chemically modified PMMA slides, silanized glass, commercial silylated glass and commercial plastic Euray trade mark slides. Immobilized and hybridized densities of 10 and 0.75 pmol/cm2, respectively, were observed for microarrays on chemically aminated PMMA. The immobilized probes were heat stable since the hybridization performance of microarrays subjected to 20 PCR heat cycles was only reduced by 4%. In conclusion, this new strategy to modify PMMA provides a robust procedure to immobilize DNA, which is a very useful substrate for fabricating single use diagnostics devices with integrated functions, like sample preparation, treatment and detection using microfabrication and microelectronic techniques.
Fixe
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INESC, Microsistemas e Nanotecnologias (INESC-MN), Rua Alves Redol, 9, 1000-029 Lisboa, Portugal.
Dufva
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Telleman
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Christensen
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D001483:Base Sequence; D015342:DNA Probes / Q000235:genetics / Q000378:metabolism; D005898:Glass; D006358:Hot Temperature; D007700:Kinetics; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods*; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism; D010969:Plastics; D016133:Polymerase Chain Reaction; D019904:Polymethyl Methacrylate / Q000737:chemistry*; D012821:Silanes
D015342:DNA Probes; D009841:Oligonucleotides; D010969:Plastics; D012821:Silanes; D019904:Polymethyl Methacrylate