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.
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Citations
64 Claims
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1-35. -35. (canceled)
- 36. A disposable unit for conducting a thermal cycling reaction, the unit comprising a thermally conducting layer and a facing layer having one or more reagent wells defined therebetween, wherein the one or more reagent wells are fed by one or more channels and each of the one more reagent wells has a single opening onto one of the one or more channels.
- 61. A kit for conducting a polymerase chain reaction, the kit comprising a buffer system comprising a buffer of pH in excess of 8.3, and at least one disposable unit comprising a thermally conducting layer and a facing layer having one or more reagent wells of up to 1000 microns depth defined therebetween.
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63. A disposable unit for conducting a thermal cycling reaction, the unit comprising a thermally conducting layer and a facing layer having a plurality of reagent wells defined therebetween, wherein all the wells are fed by a common channel which includes a single opening to outside of the unit.
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64. A disposable unit for conducting a thermal cycling reaction, the unit comprising a thermally conducting layer and a facing layer having one or more reagent wells defined therebetween, wherein the thermally conducting layer comprises a metal.
Specification