METHODS FOR OBTAINING A SINGLE MOLECULE CONSENSUS SEQUENCE
First Claim
1. A method of obtaining a single molecule consensus sequence for a single template nucleic acid molecule, comprising:
- performing a single-molecule sequencing process on a single template molecule, wherein the single template molecule comprises;
i) a first strand having a 3′
end and a 5′
end, ii) a second strand, complementary to the first strand, and having a 3′
end and a 5′
end, and iii) a linker that links the 3′
end of the first strand to the 5′
end of the second strand,thereby generating a sequence read from said single template molecule, the read comprising a first sequence corresponding to the first strand, and a second sequence corresponding to the complementary second strand; and
comparing the first sequence and second sequence from said sequence read to obtain a single molecule consensus sequence for the single template molecule.
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Abstract
Methods obtaining a single molecule consensus sequence for a single template molecule, and for obtaining a plurality of single molecule consensus sequences for a plurality of single template molecules is provided. Template molecules having two complementary regions connected with a linker are sequenced. A single read from each template molecule can be obtained, the read containing sequence information for each of the complementary regions. Single molecule consensus sequences can be determined from these reads by comparing the sequence information of the two complementary regions.
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24 Claims
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1. A method of obtaining a single molecule consensus sequence for a single template nucleic acid molecule, comprising:
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performing a single-molecule sequencing process on a single template molecule, wherein the single template molecule comprises;
i) a first strand having a 3′
end and a 5′
end, ii) a second strand, complementary to the first strand, and having a 3′
end and a 5′
end, and iii) a linker that links the 3′
end of the first strand to the 5′
end of the second strand,thereby generating a sequence read from said single template molecule, the read comprising a first sequence corresponding to the first strand, and a second sequence corresponding to the complementary second strand; and comparing the first sequence and second sequence from said sequence read to obtain a single molecule consensus sequence for the single template molecule. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24)
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13. A method of obtaining a single molecule consensus sequence for a plurality of single template nucleic acid molecules, comprising:
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performing a plurality of single-molecule sequencing processes on a plurality of single template molecules, wherein each single template molecule comprises i) a first strand having a 3′
end and a 5′
end, ii) a second strand, complementary to the first strand, having a 3′
end and a 5′
end, and iii) a linker that links the 3′
end of the first strand to the 5′
end of the second strand,thereby generating a sequence read from each single template molecule, the read for each single template molecule comprising a first sequence corresponding to the first strand, and a second sequence corresponding to the complementary second strand of each single template molecule; and comparing the first sequence and second sequence for each said sequence read to obtain a single molecule consensus sequence for each single template molecule. - View Dependent Claims (14, 15, 16, 17, 18, 19, 20, 21, 22, 23)
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Specification