Fabrication of microarray of gel-immobilized compounds on a chip by copolymerization.
The manufacturing of microchips containing oligonucleotides and proteins immobilized within gel pads, ranging in size from 10 x 10 to 100 x 100 microns, is described. The microchips are produced by photo- or persulfate-induced copolymerization of unsaturated derivatives of biomolecules with acrylamide-bisacrylamide mixture. Oligonucleotides containing 5'-allyl or 5'-butenediol units were synthesized using standard phosphoramidite chemistry. Acryloyl residues were attached to a protein by a two-step procedure. Photopolymerization was induced by illumination of the monomer solution containing initiator with UV light through the mask. The mask was applied directly over the monomer solution or projected through a microscope. Alternatively, copolymerization was carried out in drops of aqueous solution of monomers containing ammonium persulfate. Drops with different allyl-oligonucleotides were distributed on a glass slide, and the polymerization was induced by diffusion of N,N,N',N'-tetramethylethylenediamine (TEMED) from a hexane solution that covered the aqueous drops.
Vasiliskov
A V
AV
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
Timofeev
E N
EN
Surzhikov
S A
SA
Drobyshev
A L
AL
Shick
V V
VV
Mirzabekov
A D
AD
D000178:Acrylamides / Q000737:chemistry; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004058:Diffusion; D005029:Ethylenediamines / Q000737:chemistry; D005782:Gels; D005898:Glass; D006586:Hexanes; D015336:Molecular Probe Techniques; D020411:Oligonucleotide Array Sequence Analysis; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry*; D010777:Photochemistry; D011108:Polymers / Q000737:chemistry; D012996:Solutions; D014466:Ultraviolet Rays
D000178:Acrylamides; D005029:Ethylenediamines; D005782:Gels; D006586:Hexanes; D009841:Oligonucleotides; D011108:Polymers; D012996:Solutions; C005798:N,N,N',N'-tetramethylethylenediamine