Innovative analytical methods for the characterization of attached DNA were developed for assessing the surface immobilized DNA content after amplification. This implementation resulted in an important reduction of reaction times, consumption of reagents and process automation. The derivatization process, first developed for glass slides, was then adapted to microfabricated glass channels containing integrated fluidic connections. We have characterized the derivatized surfaces for the chemical attachment of oligonucleotides and evaluate the properties relevant for the amplification process: surface density, thermal stability towards thermocycling, functionalization reproducibility and storage stability. The tricarboxylate reagent benzene-1,3,5-triacetic acid (BTA) was used to attach 5′-aminated DNA primers and templates on an aminosilanized glass surface for subsequent generation of DNA colonies by in situ solid-phase amplification.
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