title: Serial DNA immobilization in micro- and extended nanospace channels.
abstract: That focused arrays, even with a small set of ligands, provide more data than single point experiments is well established in the DNA microarray research field, but microarray technology has yet to be transferred to fused silica microchips. Fused silica microchips have several attractive features such as stability to pressure, solvents, acids and bases, and can be fabricated with minute dimensions, making them good candidates for nanofluidic research. However, due to harsh bonding conditions, DNA ligands must be immobilized after fabrication, thus preventing standard microarray spotting techniques from being used. In this paper, we provide tools for serial DNA immobilization in fused silica microchips using UV. We report the synthesis of a new UV-linker which was used to covalently couple functional DNA oligos to the inside of channels in fused silica microchips. With some simple modifications to our mask aligner, we were able to transfer OHP mask patterns, which allows the creation of basically any pattern in the channels. The functionality of the oligos was measured through the binding of fluorophore-labeled complementary target oligos. We examined parameters influencing DNA immobilization, and carry-over between spots after consecutive immobilizations inside the same channel. We also report the first successful multiple immobilizations of functional DNA oligos inside single channels of extended nanospace depth (460 nm).
authors.affiliation: Department of Applied Chemistry, School of Engineering, The University of Tokyo, 7-3-1, Hongo, Bunkyo, Tokyo 113-8656, Japan.
authors.forename: Björn
authors.initials: B
authors.lastname: Renberg
authors.forename: Kae
authors.initials: K
authors.lastname: Sato
authors.forename: Kazuma
authors.initials: K
authors.lastname: Mawatari
authors.forename: Naokazu
authors.initials: N
authors.lastname: Idota
authors.forename: Takehiko
authors.initials: T
authors.lastname: Tsukahara
authors.forename: Takehiko
authors.initials: T
authors.lastname: Kitamori
mesh_terms: D001577:Benzophenones / Q000737:chemistry; D036103:Nanotechnology / Q000379:methods*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012822:Silicon Dioxide / Q000737:chemistry; D014466:Ultraviolet Rays
chemical_list: D001577:Benzophenones; D009838:Oligodeoxyribonucleotides; C047723:benzophenone; D012822:Silicon Dioxide