METHODS AND COMPOSITIONS FOR SEQUENCING NUCLEIC ACIDS USING CHARGE
First Claim
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1. A method for sequencing nucleic acids comprising, a) incorporating one or more nucleotides into a plurality of nucleic acids in one or more reaction chambers in contact with one or more ion detectors, wherein said nucleotides comprise a 3′
- -OH blocking group, said blocking group preventing any further nucleotide incorporation and any further extension of the nucleic acids in which the nucleotide is incorporated unless removed, and b) detecting hydrogen ions released upon nucleotide incorporation by said one or more ion detectors.
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Abstract
The invention provides methods and compositions, and systems for determining the identity of nucleic acids in nucleotide sequences, including sequences with one or more homopolymer regions. The methods of the invention include improvements so as to accurately identify sequences, including the difficult homopolymer sequences that are encountered during nucleotide sequencing, such as pyrosequencing.
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16 Claims
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1. A method for sequencing nucleic acids comprising, a) incorporating one or more nucleotides into a plurality of nucleic acids in one or more reaction chambers in contact with one or more ion detectors, wherein said nucleotides comprise a 3′
- -OH blocking group, said blocking group preventing any further nucleotide incorporation and any further extension of the nucleic acids in which the nucleotide is incorporated unless removed, and b) detecting hydrogen ions released upon nucleotide incorporation by said one or more ion detectors.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A method for sequencing nucleic acids comprising, a) incorporating one or more nucleotides into a plurality of nucleic acids in one or more reaction chambers in contact with one or more charge detectors, wherein said nucleotides comprise a cleavable label and a 3′
- -OH blocking group, said blocking group preventing any further nucleotide incorporation and any further extension of the nucleic acids in which the nucleotide is incorporated unless removed, b) cleaving said cleavable label under conditions such that a charged moiety is produced, and c) detecting said charged moiety with said one or more charge detectors.
- View Dependent Claims (10, 11, 12, 13, 14, 15, 16)
Specification