| 主权项 |
1. A method of identifying a nucleotide at a defined position of a target polynudeotide, the method comprising:
a) providing an electro-switchable biosensor comprising
a primer capable of forming a hybridized polynucleotide with the target polynucleotide, wherein the primer is bound via a linker to a electro-switchable surface, andmeans for applying an alternating current voltage and determining the velocity of the hybridized polynucleotides movement in the electric field; b) contacting the primer, under conditions conducive to the formation a hybridized polynudeotide and the elongation of the primer, with the target polynudeotide and a polymerase capable of elongating the primer; c) contacting the primer with one or more nucleoside triphosphate containing identical nudeobases of one type, wherein
the nucleoside triphosphate is incorporated into the hybridized polynucleotide and the primer is elongated, if its nucleobase is complementary to a nucleobase at the corresponding position on the target polynucleotide, andthe nucleoside triphosphate is not incorporated into the hybridized polynudeotide and the primer is not elongated, if its nucleobase is not complementary to a nucleobase at the corresponding position on the target polynucleotide; d) applying an alternating current voltage to the surface and determining the velocity of the hybridized polynucleotides movement in the generated electric field, wherein a decreased velocity is indicative of the incorporation of the nucleoside triphosphate; and e) if the nucleoside triphosphate is not incorporated into the hybridized polynucleotide, repeating steps b) and c) with one or more nucleoside triphosphates containing identical nucleobases of a type different from that used in previous step(s) c), until elongation is detected, thus identifying the nucleotide at the corresponding position of the target polynucleotide. |
