Methods and compositions for incorporating nucleotides
AssigneeIsoPlexis Corporation
InventorsSteven GORDON, Jerzy OLEJNIK
Life of this patent
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Abstract
The invention provides methods and compositions, including, without limitation, algorithms, computer readable media, computer programs, apparatus, and systems for determining the identity of nucleic acids in nucleotide sequences using, for example, data obtained from sequencing by synthesis methods. The methods of the invention include correcting one or more phenomena that are encountered during nucleotide sequencing, such as using sequencing by synthesis methods. These phenomena include, without limitation, sequence lead, sequence lag, spectral crosstalk, and noise resulting from variations in illumination and/or filter responses.
Claims18
A method for correcting lead or lag effects of fluorescence signals during nucleotide sequencing, comprising:
- (a) sequencing one or more nucleic acid molecules utilizing labeled nucleotides, each labeled nucleotide having at least one detectable fluorescent label and incorporated into a corresponding nucleic acid molecule of the one or more nucleic acid molecules;
- (b) acquiring data comprising first fluorescence intensities corresponding to the labeled nucleotides at a first nucleic acid position of a nucleotide sequence;
- and
- (c) correcting a lead or lag effect on the first fluorescence intensities by subtracting from the first fluorescence intensities:
- a contribution from second fluorescent intensities to the first fluorescence intensities, the second fluorescent intensities corresponding to one or more nucleic acid positions subsequent to the first nucleic acid position;
- a contribution from third fluorescent intensities to the first fluorescence intensities, the third fluorescent intensities corresponding to one or more nucleic acid positions prior to the first nucleic acid position;
- or
- both.
Dependencies are inferred from the claim text.