METHOD FOR PREPARING DNA-CHIPS Russian patent published in 2004 - IPC

FIELD: molecular biology, bioorganic chemistry, biochemistry.

SUBSTANCE: invention can be used for preparing DNA-chips. For preparing chip synthesis of conjugate of polyamine-oligonucleotide (PA-oligo) is carried out preliminary by standard procedure. For this aim oligonucleotide comprising 5'- or 3'-terminal phosphate group is activated by pair triphenylphosphine-dipyridyldisulfide in the presence of a nucleophilic catalyst in absolute organic solvent. Formed active derivative is incubated with polyamine in suitable solvent (dimethylsulfoxide, dimethylformamide or water). As a nucleophilic catalyst methylimidazole, dimethylaminopyridine or dimethylaminopyridine N-oxide is used. As a polyamine different polyamines containing from 3 to 1000 amino-group in molecule are used. Prepared solution of conjugate PA-oligo is diluted to the required concentration = 1 x 10^-7 - 1 x 10^-4 M, NaOH is added to the final concentration 0.01-0.001 M and prepared reaction mixture is applied on solid backing as series of points and dried in air followed by washing out from unbound components. As a solid backing glass, silicon, caprone, nitrocellulose, paper, plastic plates, metal, ceramics, polymers of different nature are used. Method exhibits the more simplicity in performing and allows preparing DNA-chips with the required surface density of immobilized oligonucleotides.

EFFECT: improved preparing method.

3 cl, 7 dwg, 6 ex