Fixe F, Dufva M, Telleman P, Christensen C B V
INESC, Microsistemas e Nanotecnologias (INESC-MN), Rua Alves Redol, 9, 1000-029 Lisboa, Portugal.
Nucleic Acids Res. 2004 Jan 12;32(1):e9. doi: 10.1093/nar/gng157.
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.
开发了一种化学方法来使聚甲基丙烯酸甲酯（PMMA）基板功能化。PMMA与六亚甲基二胺反应，生成用于在微阵列中固定DNA的胺化表面。伯氨基的密度为0.29 nmol/cm²。通过固定DNA探针并与互补DNA链杂交，证实了这些伯胺的可用性。化学胺化PMMA载玻片的杂交信号和杂交效率与通过不同化学修饰的PMMA载玻片、硅烷化玻璃、商用硅烷化玻璃和商用塑料Euray商标载玻片获得的杂交信号和杂交效率相当。在化学胺化的PMMA上，微阵列的固定密度和杂交密度分别观察到为10和0.75 pmol/cm²。固定的探针具有热稳定性，因为经过20个PCR热循环的微阵列的杂交性能仅降低了4%。总之，这种修饰PMMA的新策略提供了一种固定DNA的稳健方法，PMMA是一种非常有用的基板，可用于制造具有集成功能的一次性诊断设备，如使用微加工和微电子技术进行样品制备、处理和检测。
