Intermittent detection during analytical reactions
First Claim
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1. A method of performing a nucleic acid sequencing reaction, comprising:
- a) preparing a reaction mixture comprising components of the nucleic acid sequencing reaction including a polynucleotide to be sequenced;
b) initiating the nucleic acid sequencing reaction in the reaction mixture to begin progression of the nucleic acid sequencing reaction, wherein the nucleic acid sequencing reaction is initiated during a non-illuminated period, and wherein the reaction mixture comprises an enzyme selected from the group consisting of;
a polymerase, a ligase, a ribosome, a nuclease, and a kinase; and
c) maintaining conditions that allow the nucleic acid sequencing reaction to proceed while subjecting the reaction mixture to at least one illuminated period following the non-illuminated period, wherein illumination data is collected during the at least one illuminated period, thereby performing the nucleic acid sequencing reaction.
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Abstract
Methods, devices, and systems for performing intermittent detection during analytical reactions are provided. Such methods facilitate collection of reaction data from disparate reaction times. Further, such methods are useful for reducing photo-induced damage of one or more reactants in an illuminated analytical reaction at a given reaction time. In preferred embodiments, the reaction mixture is subjected to at least one illuminated and non-illuminated period and allowed to proceed such that the time in which the reaction mixture is illuminated is less than a photo-induced damage threshold period.
131 Citations
20 Claims
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1. A method of performing a nucleic acid sequencing reaction, comprising:
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a) preparing a reaction mixture comprising components of the nucleic acid sequencing reaction including a polynucleotide to be sequenced; b) initiating the nucleic acid sequencing reaction in the reaction mixture to begin progression of the nucleic acid sequencing reaction, wherein the nucleic acid sequencing reaction is initiated during a non-illuminated period, and wherein the reaction mixture comprises an enzyme selected from the group consisting of;
a polymerase, a ligase, a ribosome, a nuclease, and a kinase; andc) maintaining conditions that allow the nucleic acid sequencing reaction to proceed while subjecting the reaction mixture to at least one illuminated period following the non-illuminated period, wherein illumination data is collected during the at least one illuminated period, thereby performing the nucleic acid sequencing reaction. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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20. A method of performing a nucleic acid sequencing reaction, comprising:
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a) preparing a reaction mixture comprising components of the nucleic acid sequencing reaction including a polynucleotide to be sequenced; b) initiating the nucleic acid sequencing reaction in the reaction mixture to begin progression of the nucleic acid sequencing reaction, wherein the nucleic acid sequencing reaction is initiated during a non-illuminated period; and c) maintaining conditions that allow the nucleic acid sequencing reaction to proceed while subjecting the reaction mixture to multiple illuminated periods following the non-illuminated period, wherein illumination data is collected during the multiple illuminated periods to generate a plurality of noncontiguous sequence reads from the single nucleic acid template, thereby performing the nucleic acid sequencing reaction.
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Specification