Methods and devices for sequencing nucleic acids in smaller batches
AssigneeIsoPlexis Corporation
InventorsSteven Gordon, Thomas Hagerott, Edmund Golaski, Jerzy Olejnik
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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. A plurality of smaller flow cells is employed, each with a relatively small area to be imaged, in order to provide greater flexibility and efficiency.
Claims34
A method for identifying incorporation of a nucleotide analogue in a nucleic acid molecule during cycles of nucleotide incorporation via sequencing by synthesis of a target nucleic acid molecule, comprising:
- providing:
- I. at least one flow cell comprising an array of spots;
- II. a moveable support, configured to move said at least one flow cell;
- III. a reagent delivery mechanism in fluidic communication with said at least one flow cell;
- IV. a plurality of reagent reservoirs configured to supply reagents including a plurality of nucleotide analogues comprising a detectable fluorescent label;
- and
- V. an imaging platform configured to detect nucleotide analogues;
- moving said at least one flow cell from an initial position to an imaging position aligned with said imaging platform;
- performing a plurality of cycles of nucleotide incorporation, wherein performing an individual cycle of nucleotide incorporation comprises:
- (a) incorporating at least one nucleotide analogue from the reagents into a nucleic acid molecule complementary to the target nucleic acid molecule associated with a spot of the array of spots of the at least one flow cell;
- (b) washing non-incorporated nucleotide analogues from the at least one flow cell;
- (c) detecting a first fluorescence intensity of a detectable fluorescent label from the spot comprising the complementary nucleic acid molecule;
- and
- (d) detecting at least a second fluorescence intensity of a detectable fluorescent label from the spot comprising the complementary nucleic acid molecule;
- and
- identifying the incorporation of the nucleotide analogue in the complementary nucleic acid molecule using the detected first fluorescence intensity and the second fluorescence intensity.
Dependencies are inferred from the claim text.