Nucleic acid detection methods using universal priming

Details Nucleic acid detection methods using universal priming Nucleic acid detection methods using universal priming

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01)

Patent

CA 2399733

A method of determining the identification of nucleotides at detection positions in target sequences comprising: a) providing a plurality of probe sets, wherein each set comprises: i) a first ligation probe comprising an upstream universal priming site and a first target-specific sequence; and ii) a second ligation probe comprising a downstream universal priming site and a second target-specific sequence; wherein either the first or second target-specific sequence comprises a base at an interrogation position, and wherein at least one of said first and second ligation probes comprises an adapter sequence complementary to a capture probe on an array; b) hybridizing said plurality of probe sets to target sequences immobilized on a support or supports, said target sequences each comprising a target domain comprising a detection position and a second target domain adjacent to said detection position, wherein a first ligation probe comprising a sequence specific for the target hybridizes to one target domain and a second ligation probe comprising a sequence specific for the target hybridizes to the other target domain, and wherein if said base at said interrogation position is perfectly complementary to a nucleotide at said detection position, a ligation complex is formed; c) providing a ligase that ligates said first and second ligation probes to form ligated probes; d) amplifying said ligated probes to generate a plurality of amplicons; e) contacting said amplicons with an array comprising capture probes on a solid support; and f) determining the nucleotide at each of said detection positions.

L'invention concerne des analyses sensibles et précises pour génotypage avec une amplification minimale propre à une cible ou en son absence.

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