Reaction system for performing in the amplification of nucleic acids
First Claim
1. A method of carrying out an amplification reaction, said method comprising supplying to a reagent well in a disposable unit, a reaction mixture comprising:
- (a) a sample which contains or is suspected of containing a target nucleic acid sequence;
(b) primers, nucleotides and enzymes required to effect said amplification reaction; and
(c) a buffer system, wherein the pH of said buffer system is from 8.7-9.0, and subjecting the disposable unit to thermal cycling conditions such that any target nucleic acid present within the sample is amplified;
wherein the disposable unit comprises a thermally conducting layer and a facing layer having one or more reagent wells up to 1000 microns in depth defined therebetween, and wherein said reaction mixture further comprises a detergent and/or a blocking agent.
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Accused Products
Abstract
A method of carrying out an amplification reaction, said method comprising supplying to a well in a disposable unit (a) a sample which contains or is suspected of containing a target nucleic acid sequence (b) primers, nucleotides and enzymes required to effect said amplification reaction and (c) a buffer system, and subjecting the unit to thermal cycling conditions such that any target nucleic acid present within the sample is amplified; wherein the disposable unit comprises a thermally conducting layer and a facing layer having one or more reagent wells of up to 1000 microns in depth defined therebetween; and the reaction mixture comprises at least one of the following: A) a buffer system wherein the pH is above 8.3; B) a detergent; and/or C) a blocking agent. Apparatus for effecting the method as well as disposable units for use in the method are described. The method is particularly suitable for rapid PCR reactions.
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Citations
26 Claims
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1. A method of carrying out an amplification reaction, said method comprising supplying to a reagent well in a disposable unit, a reaction mixture comprising:
- (a) a sample which contains or is suspected of containing a target nucleic acid sequence;
(b) primers, nucleotides and enzymes required to effect said amplification reaction; and
(c) a buffer system, wherein the pH of said buffer system is from 8.7-9.0, and subjecting the disposable unit to thermal cycling conditions such that any target nucleic acid present within the sample is amplified;
wherein the disposable unit comprises a thermally conducting layer and a facing layer having one or more reagent wells up to 1000 microns in depth defined therebetween, and wherein said reaction mixture further comprises a detergent and/or a blocking agent. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
- (a) a sample which contains or is suspected of containing a target nucleic acid sequence;
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2. A method of carrying out an amplification reaction, said method comprising supplying to a reagent well in a disposable unit, a reaction mixture comprising:
- (a) a sample which contains or is suspected of containing a target nucleic acid sequence;
(b) primers, nucleotides and enzymes required to effect said amplification reaction; and
(c) a buffer system, wherein the pH of said buffer system is at or above 18.8, and subjecting the disposable unit to thermal cycling conditions such that any target nucleic acid present within the sample is amplified;
wherein the disposable unit comprises a thermally conducting layer and a facing layer having one or more reagent wells up to 1000 microns in depth defined therebetween, and wherein said reaction mixture further comprises a detergent and/or a blocking agent.
- (a) a sample which contains or is suspected of containing a target nucleic acid sequence;
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25. A method of filling a disposable unit for conducting thermal cycling reactions with a liquid, comprising:
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using air pressure to force liquid into the disposable unit; wherein the disposable unit for conducting thermal cycling reactions comprises a thermally conducting layer and a facing layer having a plurality of reagent wells defined therebetween;
wherein all the reagent wells are fed by a common channel which includes a single opening to the outside of the disposable unit;and wherein the method comprises placing the disposable unit and said liquid in a vacuum chamber, reducing pressure in said vacuum chamber such that air is evacuated from the disposable unit, immersing at least the single opening of said unit in said liquid, and increasing pressure in said vacuum chamber such that liquid is forced to enter the disposable unit through the single opening. - View Dependent Claims (26)
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Specification