公开(公告)号：US11920192B2

公开(公告)日：2024-03-05

申请号：US16613593

申请日：2018-05-15

Applicant: LIGHTCAST DISCOVERY LTD

Inventor： Barnaby Balmforth ,  Cameron Alexander Frayling ,  Mark Dethlefsen

IPC: C12Q1/6869 ,  C12Q1/6823

CPC classification number: C12Q1/6869 ,  C12Q1/6823 ,  C12Q2521/101 ,  C12Q2521/301 ,  C12Q2521/319 ,  C12Q2525/125 ,  C12Q2525/307 ,  C12Q2537/149 ,  C12Q2563/107 ,  C12Q2563/159

Abstract: 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).