NDLI: Alternative Nucleotide Flows in Sequencing-by-synthesis Methods

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Alternative Nucleotide Flows in Sequencing-by-synthesis Methods

Content Provider	The Lens
Abstract	A method of sequencing a polynucleotide strand can include providing the polynucleotide strand with a primer annealed thereto and a polymerase operably bound to the polynucleotide strand; successively exposing the polynucleotide strand to the flow of four different dNTPs according to a first predetermined ordering; and successively exposing the polynucleotide strand to the flow of four different dNTPs according to a second predetermined ordering, wherein the second predetermined ordering is different from the first predetermined ordering.
Related Links	https://www.lens.org/images/patent/081-453-313-630-899/pdf/US_2014_A1_20140031238_081-453-313-630-899.pdf
Language	English
Publisher Date	2019-08-27
Access Restriction	Open
Content Type	Text
Resource Type	Patent
Jurisdiction	United States of America
Date Applied	2017-01-12
Agent	Jones Robb, P.l.l.c.
Applicant	Life Technologies Corp
Application No.	201715404735
Claim	A method for sequencing a template polynucleotide strand comprising: (a) disposing a plurality of template polynucleotide strands into a plurality of reaction chambers, such that each reaction chamber has a template polynucleotide strand of the plurality having a sequencing primer hybridized thereto and a polymerase operably bound thereto; (b) introducing a known nucleoside triphosphate into each reaction chamber according to a predetermined ordering of dNTP reagent flows; (c) detecting sequential incorporation at the 3′ end of the sequencing primer of one or more nucleoside triphosphates if the known nucleoside triphosphate is complementary to corresponding nucleotides in the template polynucleotide strands; (d) washing away unincorporated nucleoside triphosphates from the reaction chamber; and (e) repeating steps (b) through (d) until a desired template length of the plurality of template polynucleotide strands is sequenced; wherein the predetermined ordering of dNTP reagent flows 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, and 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, and wherein repetition of the alternate ordering successively occurs at least once to sequence the desired template length. The method of claim 1 , wherein repetition of the alternate ordering successively occurs at least twice. The method of claim 1 , wherein the predetermined ordering of dNTP reagent flows comprises one of 20, 24, 28 and 32 dNTP reagent flows. A method of sequencing a polynucleotide strand, comprising: providing the polynucleotide strand with a primer annealed thereto and a polymerase operably bound to the polynucleotide strand; successively exposing the polynucleotide strand to a first series of flows of four different dNTPs according to a first predetermined ordering; and successively exposing the polynucleotide strand to a second series of flows of four different dNTPs according to a second predetermined ordering, wherein: the second predetermined ordering is different from the first predetermined ordering, a same number of flows of each of the four different dNTPs occurs in the first series of flows and the second series of flows, and repetition of the first and second predetermined orderings successively occurs at least once to sequence a desired template length, wherein one of the first or second predetermined orderings comprises 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. The method of claim 4 , wherein the second predetermined ordering comprises an alternate ordering which is not a continuous repeat of the first predetermined ordering of the four different dNTPs. The method of claim 4 , further comprising successively exposing the polynucleotide strand to a third series of flows of four different dNTPs according to a third predetermined ordering, wherein the third predetermined ordering comprises an alternate ordering which is not a continuous repeat of the first predetermined ordering of the four different dNTPs or of the second predetermined ordering of the four different dNTPs. The method of claim 4 , further comprising successively exposing the polynucleotide strand to a third series of flows of four different dNTPs according to the first predetermined ordering after successively exposing the polynucleotide strand to the second series of flows of four different dNTPs according to the second predetermined ordering. The method of claim 4 , wherein repetition of each of the first and second predetermined orderings successively occurs at least twice. The method of claim 4 , wherein each of the first and second predetermined orderings of dNTP reagent flows comprises one of 20, 24, 28 and 32 dNTP reagent flows.
CPC Classification	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 Chemical Or Physical Laboratory Apparatus For General Use
Examiner	Neil P Hammell
Extended Family	153-148-415-925-62X 081-453-313-630-899 018-775-392-790-352 153-415-265-035-843 171-679-813-956-128 127-173-701-563-801 041-129-488-182-068 107-762-382-413-397 081-050-024-703-768 046-722-502-639-634 138-514-766-947-023 097-441-733-975-554 151-829-635-602-794 063-920-121-800-168 192-903-349-554-656 011-377-015-913-021 033-543-655-259-821 171-734-174-498-536 002-689-398-892-171
Patent ID	10392660
Inventor/Author	Schultz Jonathan Davidson John
IPC	B01L3/00 C12Q1/6869 C12Q1/6874
Status	Active
Owner	Life Technologies Corporation
Simple Family	153-148-415-925-62X 081-453-313-630-899 018-775-392-790-352 153-415-265-035-843 171-679-813-956-128 127-173-701-563-801 041-129-488-182-068 107-762-382-413-397 081-050-024-703-768 046-722-502-639-634 138-514-766-947-023 097-441-733-975-554 151-829-635-602-794 192-903-349-554-656 063-920-121-800-168 011-377-015-913-021 033-543-655-259-821 171-734-174-498-536 002-689-398-892-171
CPC (with Group)	C12Q1/6874 B01L3/5027 C12Q1/6869
Issuing Authority	United States Patent and Trademark Office (USPTO)
Kind	Patent/New European patent specification (amended specification after opposition procedure)

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