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AssigneeLightcast Discovery Ltd
InventorsBarnaby Balmforth, Cameron Alexander Frayling, Mark Dethlefsen
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A method of sequencing a nucleic acid comprising (1) generating a stream of single nucleoside triphosphates by progressive enzymatic digestion of the nucleic acid; (2) producing at least one oligonucleotide used probe by reacting, in the presence of a polymerase, at least one of the single nucleoside triphosphates with a corresponding biological probe comprising (a) a first single-stranded oligonucleotide including an exonuclease blocking-site, a restriction endonuclease recognition-site located on the 5′ side of the blocking-site and including a single nucleotide capture-site e, and at least one fluorophore region and (b) a second and optionally a third single-stranded oligonucleotide each separate from the first oligonucleotide; (3) cleaving the first oligonucleotide strand of the used probe at the recognition-site with a restriction endonuclease; (4) digesting the first oligonucleotide component with an enzyme to yield fluorophores in a detectable state and (5) detecting the fluorophores released in step (4).
A multi-component biological probe for capturing a single nucleoside triphosphate, the probe comprising a multiplicity of different first single-stranded oligonucleotides, wherein (a) each of the first single-stranded oligonucleotides include an exonuclease blocking-site, a restriction endonuclease recognition-site located on the 5′ side of the blocking-site and including a single nucleotide capture-site, and at least one fluorophore region located on the 5′ side of the recognition-site and arranged so as to render the fluorophore(s) quenched and (b) separate second and optionally separate third single-stranded oligonucleotides capable of hybridising to complementary regions on the first oligonucleotide either side of the capture-site wherein the multiplicity of different first oligonucleotide types differs only in the nucleobase characteristic of the capture-site and the fluorophore(s);
Dependencies are inferred from the claim text.