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Alternative Nucleotide Flows in Sequencing-by-synthesis Methods
| Content Provider | The Lens |
|---|---|
| Abstract | A method for sequencing a polynucleotide strand by using sequencing-by-synthesis techniques. To address the problem of incomplete extension (IE) and/or carry forward (CF) errors that can occur in sequencing-by-synthesis reactions, an alternative flow ordering of dNTPs is used. In contrast to conventional flow orderings, the dNTPs are flowed in an ordering that is not a continuous repeat of an ordering of the four different dNTPs. This alternate flow ordering may reduce the loss of phasic synchrony in the population of template polynucleotide strands that result from IE and/or CF errors. |
| Related Links | https://www.lens.org/lens/patent/011-377-015-913-021/frontpage |
| Language | English |
| Publisher Date | 2017-03-28 |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Patent |
| Jurisdiction | United States of America |
| Date Applied | 2012-11-29 |
| Agent | Jones Robb, Pllc |
| Applicant | Life Technologies Corp |
| Application No. | 201213689252 |
| Claim | An apparatus for sequencing a polynucleotide strand, comprising: a flow chamber for receiving flows of different dNTP reagents; multiple reservoirs that each contain a different dNTP reagent; flow paths from each of the reservoirs to the flow chamber; and a fluidics controller that controls the flow from the reservoirs to the flow chamber, wherein the fluidics controller is programmed to successively provide flow from the multiple reservoirs to the flow chamber according to a predetermined ordering, wherein the predetermined ordering comprises an alternate ordering which is not a continuous and ordered repeat of four different dNTP reagent flows, the alternate ordering comprising a same number of flows for each of four different dNTP reagent flows, a flow of a nucleotide N, which is followed directly by a flow of a different nucleotide W, which is followed directly by a flow of the same nucleotide N, wherein the N-W-N flow is further followed directly by a flow of a nucleotide Z that is different from N and W, and the alternate ordering being successively repeated at least once. The apparatus of claim 1 , wherein the alternate ordering is not a continuous repeat of an order of a first four dNTP reagent flows in the predetermined ordering. The apparatus of claim 1 , further comprising a flow cell loaded into the flow chamber, wherein the flow cell comprises a microwell array that contains multiple copies of the polynucleotide strand with a primer annealed thereto. The apparatus of claim 3 , wherein the flow cell further comprises a chemFET sensor array for detecting the reaction of the dNTP reagent with the contents of the microwell array. The apparatus of claim 3 , wherein the polynucleotide strands are attached to beads that are contained in the microwells of the microwell array. The apparatus of claim 1 , wherein the predetermined ordering consists of only the alternate ordering. The apparatus of claim 1 , wherein the predetermined ordering further comprises a continuous and ordered flow of the four different dNTP reagent flows as follows: dATP, then dCTP, then dGTP, and then dTTP. The apparatus of claim 1 , wherein a first four dNTP reagent flows of the continuous and ordered repeat of the four different reagent flows comprises: dATP, then dCTP, then dGTP, and then dTTP. The apparatus of claim 1 , wherein the predetermined ordering includes a first portion that comprises the alternate ordering and a second portion that does not comprise the alternate ordering. The apparatus of claim 1 , wherein in the predetermined ordering, the flow of one of the four dNTP reagent flows is always followed by a flow of a different one of the four dNTP reagent flows. The apparatus of claim 1 , wherein the predetermined ordering further comprises a continuous and ordered repeat of the four different dNTP reagent flows and is a permutation of a flow ordering of: dATP, then dCTP, then dGTP, and then dTTP. The apparatus of claim 1 , wherein the alternate ordering is repeated two or more times. An apparatus for sequencing a polynucleotide strand, comprising: a flow chamber for receiving flows of different dNTP reagents; multiple reservoirs that each contain a different dNTP reagent; flow paths from each of the reservoirs to the flow chamber; and a fluidics controller that controls the flow from the reservoirs to the flow chamber, wherein the fluidics controller is programmed to successively provide flow from the multiple reservoirs to the flow chamber according to a predetermined ordering, wherein the predetermined ordering comprises an alternate ordering which is not a continuous and ordered repeat of four different dNTP reagent flows, the alternate ordering comprising a flow of one kind of dNTP reagent always followed by a flow of a different kind of dNTP reagent, at least one flow of each kind of dNTP reagent followed by a flow of the same kind of dNTP reagent after a single intervening flow of a different kind of dNTP reagent, a same number of flows of each of the four kinds of dNTP reagent flows in the alternate ordering, and wherein repetition of the alternate ordering successively occurs at least once to sequence a desired template length. The apparatus of claim 13 , wherein the alternate ordering is repeated two or more times. The apparatus of claim 1 , wherein the plurality of reagent flows in the alternate ordering comprises one of 8, 12, 16, 20, 24, 28 and 32 dNTP reagent flows. The apparatus of claim 13 , wherein the plurality of reagent flows in the alternate ordering comprises one of 8, 12, 16, 20, 24, 28 and 32 dNTP reagent flows. An apparatus for sequencing a polynucleotide strand, comprising: a flow chamber for receiving flows of different dNTP reagents; multiple reservoirs that each contain a different dNTP reagent; flow paths from each of the reservoirs to the flow chamber; and a fluidics controller that controls the flow from the reservoirs to the flow chamber, wherein the fluidics controller is programmed to successively provide flow from the multiple reservoirs to the flow chamber according to a predetermined ordering, wherein the predetermined ordering consists of only an alternate ordering which is not a continuous and ordered repeat of four different dNTP reagent flows, the alternate ordering comprising a same number of flows for each of four different dNTP reagent flows, a flow of a nucleotide N, which is followed directly by a flow of a different nucleotide W, which is followed directly by a flow of the same nucleotide N, wherein the N-W-N flow is further followed directly by a flow of a nucleotide Z that is different from N and W, and the alternate ordering being repeated at least once. |
| CPC Classification | Chemical Or Physical Laboratory Apparatus For General Use MEASURING OR TESTING PROCESSES INVOLVING ENZYMES; NUCLEIC ACIDS OR MICROORGANISMS ;COMPOSITIONS OR TEST PAPERS THEREFOR;PROCESSES OF PREPARING SUCH COMPOSITIONS;CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES |
| Examiner | Neil P Hammell |
| Extended Family | 171-734-174-498-536 081-453-313-630-899 018-775-392-790-352 153-415-265-035-843 107-604-770-612-773 171-679-813-956-128 041-129-488-182-068 127-173-701-563-801 107-762-382-413-397 081-050-024-703-768 046-722-502-639-634 138-514-766-947-023 097-441-733-975-554 151-829-635-602-794 011-377-015-913-021 192-903-349-554-656 063-920-121-800-168 033-543-655-259-821 002-689-398-892-171 153-148-415-925-62X |
| Patent ID | 9605308 |
| Inventor/Author | Schultz Jonathan Davidson John |
| IPC | C12Q1/68 B01L3/00 |
| Status | Active |
| Owner | Life Technologies Corporation |
| Simple Family | 171-734-174-498-536 081-453-313-630-899 018-775-392-790-352 153-415-265-035-843 107-604-770-612-773 171-679-813-956-128 041-129-488-182-068 127-173-701-563-801 107-762-382-413-397 081-050-024-703-768 046-722-502-639-634 138-514-766-947-023 097-441-733-975-554 151-829-635-602-794 063-920-121-800-168 192-903-349-554-656 011-377-015-913-021 033-543-655-259-821 002-689-398-892-171 153-148-415-925-62X |
| CPC (with Group) | B01L3/5027 C12Q1/6874 C12Q1/6869 |
| Issuing Authority | United States Patent and Trademark Office (USPTO) |
| Kind | Patent/New European patent specification (amended specification after opposition procedure) |
