Direct ATP release sequencing
AssigneeAgilent Technologies, Inc.
InventorJeffrey R. Sampson
Life of this patent
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Abstract
A method for sequencing a nucleic acid is provided. In certain embodiments, the method includes contacting a nucleic acid duplex comprising a nucleic acid template and a primer annealed to the template with a reagent mix under primer extension conditions to produce an extended primer and ATP. The reagent mix may contain an adenosine-2′-deoxynucleoside tetraphosphate moiety and a polymerase. The method further includes detecting the produced ATP. Also provided are adenosine-2′-deoxynucleoside tetraphosphate moieties that find use in the subject methods. Also provided are kits containing the adenosine-2′-deoxynucleoside tetraphosphate moieties for use in the subject methods.
Claims20
A method comprising:
- (a) contacting a nucleic acid duplex comprising a nucleic acid template and a primer annealed to said nucleic acid template with a reagent mix comprising an adenosine-2′-deoxynucleoside tetraphosphate moiety and a polymerase under primer extension conditions to produce an extended primer and adenosine triphosphate (ATP);
- and
- (b) detecting said ATP.
A method of sequencing a nucleic acid comprising:
- (a) separately contacting a nucleic acid duplex comprising a nucleic acid template and a primer annealed to said nucleic acid template with:
- (i) adenosine-2′-deoxycytidine tetraphosphate;
- (ii) adenosine-2′-deoxyguanosine tetraphosphate;
- (iii) adenosine-2′-deoxyadenosine tetraphosphate;
- or
- (iv) adenosine-thymidine tetraphosphate;
- and
- a polymerase under primer extension conditions to produce a plurality of reactions;
- (b) detecting which of said reactions produces ATP, wherein ATP production indicates the nucleotide added to said primer.
A method of sequencing a nucleic acid comprising:
- (a) sequentially contacting a nucleic acid duplex comprising a nucleic acid template and a primer annealed to said nucleic acid template with, in any order:
- (i) adenosine-2′-deoxycytidine tetraphosphate;
- (ii) adenosine-2′-deoxyguanosine tetraphosphate;
- (iii) adenosine-2′-deoxyadenosine tetraphosphate;
- and
- (iv) adenosine-thymidine tetraphosphate;
- and
- a polymerase under primer extension conditions;
- and
- (b) detecting which of said reactions produces ATP, wherein ATP production indicates the nucleotide added to said primer.
A kit comprising:
- (i) adenosine-2′-deoxycytidine tetraphosphate;
- (ii) adenosine-2′-deoxyguanosine tetraphosphate;
- (iii) adenosine-2′-deoxyadenosine tetraphosphate;
- (iv) adenosine-thymidine tetraphosphate;
- and
- (v) a polymerase.
Dependencies are inferred from the claim text.