Methods and compositions for sequencing nucleic acid using charge
First Claim
Patent Images
1. A method for sequencing nucleic acids comprising, a) incorporating a nucleotide into one or more nucleic acids of a plurality of nucleic acids in one or more reaction chambers in contact with one or more ion detectors, wherein said nucleotides comprise a removable 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, wherein said nucleic acids comprise incorporated nucleotides in a chain and a portion of the sequence of said nucleic acids comprises consecutive identical bases of the formula Xn, where X is any base and n is a whole number between 3 and 10, b) detecting hydrogen ions released upon said nucleotide incorporation by said one or more ion detectors, said hydrogen ions coming from the 3′
-OH group of an already incorporated nucleotide in the chain when the new nucleotide is incorporated, and c) sequencing the nucleic acids by determining the identity of the incorporated nucleotide.
6 Assignments
0 Petitions
Accused Products
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.
-
Citations
7 Claims
-
1. A method for sequencing nucleic acids comprising, a) incorporating a nucleotide into one or more nucleic acids of a plurality of nucleic acids in one or more reaction chambers in contact with one or more ion detectors, wherein said nucleotides comprise a removable 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, wherein said nucleic acids comprise incorporated nucleotides in a chain and a portion of the sequence of said nucleic acids comprises consecutive identical bases of the formula Xn, where X is any base and n is a whole number between 3 and 10, b) detecting hydrogen ions released upon said nucleotide incorporation by said one or more ion detectors, said hydrogen ions coming from the 3′
-OH group of an already incorporated nucleotide in the chain when the new nucleotide is incorporated, and c) sequencing the nucleic acids by determining the identity of the incorporated nucleotide. - View Dependent Claims (2, 3, 4)
- -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, wherein said nucleic acids comprise incorporated nucleotides in a chain and a portion of the sequence of said nucleic acids comprises consecutive identical bases of the formula Xn, where X is any base and n is a whole number between 3 and 10, b) detecting hydrogen ions released upon said nucleotide incorporation by said one or more ion detectors, said hydrogen ions coming from the 3′
-
5. A method for sequencing nucleic acids comprising, a) introducing nucleotides in one or more reaction chambers in contact with one or more ion detectors, wherein said reaction chambers comprise nucleic acid having a sequence, a portion of which comprising consecutive identical bases of the formula Xn, where X is any base and n is a whole number between 3 and 10, and wherein said nucleotides comprise a 3′
- -OH blocking group comprising a removable chemical moiety, 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) incorporating a first nucleotide from said nucleotides into said nucleic acid with a polymerase, c) detecting hydrogen ions released upon said first nucleotide incorporation by said one or more ion detectors, d) removing said 3′
-OH blocking group, e) incorporating a second nucleotide from said nucleotides into said nucleic acid with a polymerase, and f) sequencing the nucleic acids by determining the identity of the incorporated nucleotides. - View Dependent Claims (6, 7)
- -OH blocking group comprising a removable chemical moiety, 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) incorporating a first nucleotide from said nucleotides into said nucleic acid with a polymerase, c) detecting hydrogen ions released upon said first nucleotide incorporation by said one or more ion detectors, d) removing said 3′
Specification