Methods and compositions for nucleic acid sequencing
First Claim
Patent Images
1. A method for determining sequences of polynucleotides, the method comprising:
- (a) performing a sequencing reaction that comprises repeated cycles of;
(i) incorporating nucleotide conjugates into a plurality of polynucleotides to produce extended polynucleotides,(ii) detecting a first collection of signals from the extended polynucleotides, wherein the first collection of signals comprises signals from a first label attached to a first type of the nucleotide conjugates and a second label attached to a second type of the nucleotide conjugates,(iii) adding a third label to a third type of the nucleotide conjugates comprising a hapten that are incorporated into the extended polynucleotides, thereby producing modified polynucleotides; and
(iv) detecting a second collection of signals from the modified polynucleotides, wherein the second collection of signals comprises signals from the third label added to the third type of the nucleotide conjugates; and
(b) determining incorporation of a fourth type of the nucleotide conjugates based on a detection pattern of the first, second and third types of the nucleotide conjugates into the extended polynucleotides, wherein the incorporation of the fourth type of the nucleotide conjugates is determined from a dark state in the first and second collection of signals.
3 Assignments
0 Petitions
Accused Products
Abstract
The present disclosure provides methods and systems for detecting multiple different nucleotides in a sample. In particular, the disclosure provides for detection of multiple different nucleotides in a sample utilizing fewer detection moieties than the number of nucleotides being detected and/or fewer imaging events than the number of nucleotides being detected.
-
Citations
19 Claims
-
1. A method for determining sequences of polynucleotides, the method comprising:
-
(a) performing a sequencing reaction that comprises repeated cycles of; (i) incorporating nucleotide conjugates into a plurality of polynucleotides to produce extended polynucleotides, (ii) detecting a first collection of signals from the extended polynucleotides, wherein the first collection of signals comprises signals from a first label attached to a first type of the nucleotide conjugates and a second label attached to a second type of the nucleotide conjugates, (iii) adding a third label to a third type of the nucleotide conjugates comprising a hapten that are incorporated into the extended polynucleotides, thereby producing modified polynucleotides; and (iv) detecting a second collection of signals from the modified polynucleotides, wherein the second collection of signals comprises signals from the third label added to the third type of the nucleotide conjugates; and (b) determining incorporation of a fourth type of the nucleotide conjugates based on a detection pattern of the first, second and third types of the nucleotide conjugates into the extended polynucleotides, wherein the incorporation of the fourth type of the nucleotide conjugates is determined from a dark state in the first and second collection of signals. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
-
Specification