METHOD OF POLYNUCLEOTIDE SEQUENCING AND DEVICE FOR ITS REALIZATION Russian patent published in 2003 - IPC

FIELD: genetic engineering, molecular biology, biochemistry. SUBSTANCE: method involves interaction of polynucleotide-target with polymerase immobilized on solid backing and different nucleotides under conditions providing polymerase reaction running. Incorporation of specific nucleotide which is complementary to polynucleotide-target is found by radiation measurement. Backing presents sensory chip. Both stages can be carried out with each of different nucleotides by turns up to moment when incorporation is detected followed by repeating these stages. The first stage can be carried out in the presence of all nucleotides. Nucleotides can comprise 3'-blocking group that is removed after polymerization reaction running. Blocking group can be removed by effect of pulsing monochromatic light. The first stage can involve additional addition of competitive inhibitor of polymerase. DNA-polymerase-III from E. coli or Taq- polymerase, or reverse transcriptase can be used as polymerase. Radiation can be electromagnetic one and relates to infrared region of spectrum or to radio-frequency spectrum. Device consists of optical sensory chip carrying immobilized polymerase. Chip is used in plasmon resonance on surface or in nuclear magnetic resonance of the light source. Device involves equipment for image reading and light-sensitive unit. Invention allows to accelerate polynucleotide sequencing, decrease its labor intensity and price. EFFECT: improved method of sequencing. 22 cl, 5 dwg