Reaction System for Performing in the Amplification of Nucleic Acids
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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.
23 Citations
55 Claims
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1-35. -35. (canceled)
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36. A method of carrying out a rapid amplification reaction, the method comprising supplying to a reagent well in a disposable unit:
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(a) a sample that contains or is suspected of containing a target nucleic acid sequence; (b) primers, nucleotides and enzymes required to effect the amplification reaction; (c) a buffer system; and (d) a blocking agent, and subjecting the disposable unit to amplification conditions such that 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. - View Dependent Claims (37, 38, 39, 40)
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41. A method of carrying out an amplification reaction, the method comprising supplying to a reagent well in a disposable unit:
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(a) a sample that contains or is suspected of containing a target nucleic acid sequence; (b) primers, nucleotides and enzymes required to effect the amplification reaction; and (c) a buffer system having a concentration of 30-70 mM; and (d) a blocking agent, 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 of up to 1000 microns in depth defined therebetween. - View Dependent Claims (42, 43, 44, 45, 46, 47)
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- 48. A disposable unit for conducting a thermal cycling reaction, wherein the disposable unit comprises a pouch comprising a thermally conducting layer and a facing layer having a plurality of reagent wells defined therebetween, wherein the reagent wells are fed by a common channel that includes a single opening to an outside of the pouch.
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55. A kit comprising:
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(a) a disposable unit for conducting a thermal cycling reaction; (b) a buffer system of 30 to 70 mM; and (c) a blocking agent, wherein the disposable unit comprises a pouch comprising a thermally conducting layer and a facing layer having a plurality of reagent wells defined therebetween, wherein the reagent wells are fed by a common channel that includes a single opening to an outside of the pouch.
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Specification