EDC-mediated DNA attachment to nanocrystalline CVD diamond films.
Chemical vapour deposited (CVD) diamond is a very promising material for biosensor fabrication owing both to its chemical inertness and the ability to make it electrical semiconducting that allows for connection with integrated circuits. For biosensor construction, a biochemical method to immobilize nucleic acids to a diamond surface has been developed. Nanocrystalline diamond is grown using microwave plasma-enhanced chemical vapour deposition (MPECVD). After hydrogenation of the surface, 10-undecenoic acid, an omega-unsaturated fatty acid, is tethered by 254 nm photochemical attachment. This is followed by 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide (EDC)-mediated attachment of amino (NH(2))-modified dsDNA. The functionality of the covalently bound dsDNA molecules is confirmed by fluorescence measurements, PCR and gel electrophoresis during 35 denaturation and rehybridisation steps. The linking method after the fatty acid attachment can easily be applied to other biomolecules like antibodies and enzymes.
Christiaens
P
P
Hasselt University, Institute for Materials Research and Transnationale Universiteit Limburg, School for Life Sciences, Wetenschapspark 1, B-3590 Diepenbeek, Belgium. Pieter.Christiaens@uhasselt.be
Vermeeren
V
V
Wenmackers
S
S
Daenen
M
M
Haenen
K
K
Nesládek
M
M
vandeVen
M
M
Ameloot
M
M
Michiels
L
L
Wagner
P
P
D015374:Biosensing Techniques / Q000295:instrumentation*; D004247:DNA / Q000138:chemical synthesis; D018130:Diamond; D005022:Ethyldimethylaminopropyl Carbodiimide; D005231:Fatty Acids, Unsaturated; D053758:Nanoparticles
D005231:Fatty Acids, Unsaturated; D018130:Diamond; D004247:DNA; D005022:Ethyldimethylaminopropyl Carbodiimide