Methods, systems, computer readable media, and kits for sample identification
First Claim
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1. A method for sequencing a polynucleotide, the method comprising:
- introducing, into a polynucleotide, a barcode sequence to create a polynucleotide sample, wherein the barcode sequence is a series of nucleotide bases determined by a predetermined flowspace vector corresponding to a predetermined order of nucleotide flows used for sequencing, wherein the predetermined flowspace vector is a string of characters with each character representing an expected number of nucleotide incorporations resulting from a nucleotide flow of the predetermined order of nucleotide flows;
inserting the created polynucleotide sample into a flow chamber of a sequencing instrument comprising the flow chamber, a sensor in the flow chamber, and a fluidics controller configured to control the flow of nucleotides into the flow chamber;
exposing the created polynucleotide sample in the flow chamber to the series of nucleotides in a presence of a polymerase and a primer by individually flowing the series of nucleotides into the flow chamber according to the predetermined order of nucleotide flows using the fluidics controller, wherein incorporation of nucleotides of the series of nucleotides into the created polynucleotide sample produces a synthesized polynucleotide complementary to the polynucleotide sample;
obtaining, using the sensor, a series of signals resulting from the exposing of the created polynucleotide sample to the series of nucleotides, the series of signals indicating one or more nucleotide incorporations or one or more nucleotide non-incorporations for each nucleotide flow;
outputting the series of signals as data to a computing unit in communication with the sensor, the computing unit comprising a processor and a memory configured to receive and store the data;
resolving, using the processor, the series of signals to generate a rendered flowspace string of characters, wherein each character represents a number of nucleotide incorporations resulting from a respective nucleotide flow of the predetermined order of nucleotide flows;
comparing, using the processor, the rendered flowspace string of characters to predetermined flowspace strings of characters corresponding to a set of barcode sequences comprising the introduced barcode sequence, wherein the predetermined flowspace strings of characters are codewords of an error-tolerant code, the error-tolerant code is an error-correcting code configured to correct one or more errors present in the rendered flowspace string of characters based on differences between the rendered flowspace string and the predetermined flowspace strings of characters; and
identifying, by the processor, the polynucleotide of the polynucleotide sample based on the comparison.
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Abstract
A method for sequencing a polynucleotide sample having a barcode sequence, includes: introducing a series of nucleotides to the polynucleotide sample according to a predetermined flow ordering; obtaining a series of signals resulting from the introducing of nucleotides to the polynucleotide sample; and resolving the series of signals over the barcode sequence to render a flowspace string, wherein the flowspace string is a codeword of an error-tolerant code capable of distinguishing the barcode sequence from other barcode sequences in the presence of one or more errors.
126 Citations
20 Claims
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1. A method for sequencing a polynucleotide, the method comprising:
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introducing, into a polynucleotide, a barcode sequence to create a polynucleotide sample, wherein the barcode sequence is a series of nucleotide bases determined by a predetermined flowspace vector corresponding to a predetermined order of nucleotide flows used for sequencing, wherein the predetermined flowspace vector is a string of characters with each character representing an expected number of nucleotide incorporations resulting from a nucleotide flow of the predetermined order of nucleotide flows; inserting the created polynucleotide sample into a flow chamber of a sequencing instrument comprising the flow chamber, a sensor in the flow chamber, and a fluidics controller configured to control the flow of nucleotides into the flow chamber; exposing the created polynucleotide sample in the flow chamber to the series of nucleotides in a presence of a polymerase and a primer by individually flowing the series of nucleotides into the flow chamber according to the predetermined order of nucleotide flows using the fluidics controller, wherein incorporation of nucleotides of the series of nucleotides into the created polynucleotide sample produces a synthesized polynucleotide complementary to the polynucleotide sample; obtaining, using the sensor, a series of signals resulting from the exposing of the created polynucleotide sample to the series of nucleotides, the series of signals indicating one or more nucleotide incorporations or one or more nucleotide non-incorporations for each nucleotide flow; outputting the series of signals as data to a computing unit in communication with the sensor, the computing unit comprising a processor and a memory configured to receive and store the data; resolving, using the processor, the series of signals to generate a rendered flowspace string of characters, wherein each character represents a number of nucleotide incorporations resulting from a respective nucleotide flow of the predetermined order of nucleotide flows; comparing, using the processor, the rendered flowspace string of characters to predetermined flowspace strings of characters corresponding to a set of barcode sequences comprising the introduced barcode sequence, wherein the predetermined flowspace strings of characters are codewords of an error-tolerant code, the error-tolerant code is an error-correcting code configured to correct one or more errors present in the rendered flowspace string of characters based on differences between the rendered flowspace string and the predetermined flowspace strings of characters; and identifying, by the processor, the polynucleotide of the polynucleotide sample based on the comparison. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 19)
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18. A system comprising:
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a sequencing instrument comprising; a flow chamber configured to receive a polynucleotide sample, a polymerase, and a primer, the polynucleotide sample comprising a polynucleotide incorporating a barcode sequence, wherein the barcode sequence is a series of nucleotide bases determined by a predetermined flowspace vector corresponding to a predetermined order of nucleotide flows used for sequencing, wherein the predetermined flowspace vector is a string of characters with each character representing an expected number of nucleotide incorporations resulting from a nucleotide flow of the predetermined order of nucleotide flows; a fluidics controller configured to flow nucleotides into the flow chamber to expose the polynucleotide sample to a series of nucleotides by individually flowing the series of nucleotides according to the predetermined order of nucleotide flows, and a sensor configured to obtain a series of signals resulting from the exposing of the polynucleotide sample to the series of nucleotides, and output the series of signals as data, the series of signals indicating one or more nucleotide incorporations or one or more nucleotide non-incorporations for each nucleotide flow; and a computing unit in communication with the sensor, the computing unit comprising a processor and a memory configured to receive and store the data, the processor configured to; resolve the series of signals to generate a rendered flowspace string of characters, wherein each character represents a number of nucleotide incorporations resulting from a respective nucleotide flow of the predetermined order of nucleotide flows; compare the rendered flowspace string of characters to predetermined flowspace strings of characters corresponding to a set of barcode sequences comprising the incorporated barcode sequence, wherein the predetermined flowspace strings of characters are codewords of an error-tolerant code, the error-tolerant code is an error-correcting code configured to correct one or more errors present in the rendered flowspace string of characters based on differences between the rendered flowspace string of characters and the predetermined flowspace strings of characters; and identify the polynucleotide of the polynucleotide sample based on the comparison. - View Dependent Claims (20)
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Specification