Nucleic acid amplification reaction apparatus
First Claim
1. An apparatus for performing a nucleic acid amplification reaction in a reaction mixture in a capillary tube, the nucleic acid amplification reaction including denaturation, annealing and extension processes, the apparatus comprising:
- (a) a first heat exchanger including a thermoregulating system for stabilizing the temperature of the first heat exchanger at a temperature in a range of temperatures suitable to cause the denaturation process to occur in the reaction mixture;
(b) a second heat exchanger including a thermoregulating system for stabilizing the temperature of the second heat exchanger at a temperature in a range of temperatures suitable for causing the annealing and extension processes to occur in the reaction mixture;
(c) the capillary tube being routed so as to have a first portion in thermal contact with the first heat exchanger and a second portion in thermal contact with the second heat exchanger;
(d) a valve assembly having an input port coupled to the capillary tube for receiving said reaction mixture into the tube and an output port for delivering a finished reaction product from the tube, the valve assembly allowing injection of new reaction mixture in a first position, allowing the reaction mixture to circulate in the capillary tube between the first and second heat exchangers in a second position, and allowing ejection of finished reaction product in a third position; and
(e) a pumping device fixedly disposed in one of the heat exchangers coupled to the capillary tube so as to circulate the reaction mixture in the capillary tube such that the reaction mixture passes sequentially and cyclically through the first and second heat exchangers.
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Abstract
Apparatus and method for performing a nucleic acid amplification reaction and preferably a polymerase chain reaction (PCR) in a reaction mixture in at least one capillary tube. Several different embodiments are disclosed. One embodiment cycles a sample through a capillary tube loop passing through two thermostatted fluid baths. Another embodiment has the capillary tube routed alternatingly between two heat exchangers to that the sample makes only one pass through the tube. Other embodiments maintain the heat exchangers stationary and translate the samples between them. Still further embodiments maintain the samples stationary and either automatically translate or rotate the heat exchangers past the samples contained within the capillary tubes to perform the thermal cycles necessary for the amplification reaction.
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Citations
20 Claims
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1. An apparatus for performing a nucleic acid amplification reaction in a reaction mixture in a capillary tube, the nucleic acid amplification reaction including denaturation, annealing and extension processes, the apparatus comprising:
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(a) a first heat exchanger including a thermoregulating system for stabilizing the temperature of the first heat exchanger at a temperature in a range of temperatures suitable to cause the denaturation process to occur in the reaction mixture; (b) a second heat exchanger including a thermoregulating system for stabilizing the temperature of the second heat exchanger at a temperature in a range of temperatures suitable for causing the annealing and extension processes to occur in the reaction mixture; (c) the capillary tube being routed so as to have a first portion in thermal contact with the first heat exchanger and a second portion in thermal contact with the second heat exchanger; (d) a valve assembly having an input port coupled to the capillary tube for receiving said reaction mixture into the tube and an output port for delivering a finished reaction product from the tube, the valve assembly allowing injection of new reaction mixture in a first position, allowing the reaction mixture to circulate in the capillary tube between the first and second heat exchangers in a second position, and allowing ejection of finished reaction product in a third position; and (e) a pumping device fixedly disposed in one of the heat exchangers coupled to the capillary tube so as to circulate the reaction mixture in the capillary tube such that the reaction mixture passes sequentially and cyclically through the first and second heat exchangers. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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8. An apparatus for performing a nucleic acid amplification reaction in a reaction mixture in a capillary tube, the nucleic acid amplification reaction including denaturation, annealing and extension processes, the apparatus comprising:
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(a) a first heat exchanger including a thermoregulating system for stabilizing the temperature of the first heat exchanger at a temperature in a range of temperatures suitable to cause the denaturation process to occur in the reaction mixture; (b) a second heat exchanger including a thermoregulating system for stabilizing the temperature of the second heat exchanger at a temperature in a range of temperatures suitable for causing the annealing and extension processes to occur in the reaction mixture; (c) the capillary tube being routed so as to have a first portion in thermal contact with the first heat exchanger and a second portion in thermal contact with the second heat exchanger; (d) a valve assembly having an input port coupled to the capillary tube for receiving said reaction mixture into the tube and an output port for delivering a finished reaction product from the tube, the valve assembly allowing injection of new reaction mixture in a first position, allowing the reaction mixture to circulate in the capillary tube between the first and second heat exchangers in a second position, and allowing ejection of finished reaction product in a third position; (e) a pumping device in one of the heat exchangers coupled to the capillary tube so as to circulate the reaction mixture in the capillary tube such that the reaction mixture passes sequentially and cyclically through the first and second heat exchangers; and (f) a spectrophotometric detector optically coupled to the capillary tube for generating data representing changes in the reaction mixture during nucleic acid amplification reaction. - View Dependent Claims (9, 10, 11, 12, 13)
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14. An apparatus for performing a nucleic acid amplification reaction in a reaction mixture in a capillary tube, the nucleic acid amplification reaction including denaturation, annealing and extension processes, the apparatus comprising:
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(a) a first heat exchanger including a thermoregulating system for stabilizing the temperature of the first heat exchanger at a temperature in a range of temperatures suitable to cause the denaturation process to occur in the reaction mixture; (b) a second heat exchanger including a thermoregulating system for stabilizing the temperature of the second heat exchanger at a temperature in a range of temperatures suitable for causing the annealing and extension processes to occur in the reaction mixture; (c) the capillary tube being routed so as to have a first portion in thermal contact with the first heat exchanger and a second portion in thermal contact with the second heat exchanger; (d) a valve assembly having an input port coupled to the capillary tube for receiving said reaction mixture into the tube and an output port for delivering a finished reaction product from the tube, the valve assembly allowing injection of new reaction mixture in a first position, allowing the reaction mixture to circulate in the capillary tube between the first and second heat exchangers in a second position, and allowing ejection of finished reaction product in a third position; (e) a pumping device in one of the heat exchangers coupled to the capillary tube so as to circulate the reaction mixture in the capillary tube such that the reaction mixture passes sequentially and cyclically through the first and second heat exchangers; (f) means for injecting a discontinuity adjacent to the reaction mixture in the capillary tube; and (g) a cycle counter coupled to said capillary tube for counting the passage of said discontinuity. - View Dependent Claims (15, 16, 17, 18, 19, 20)
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Specification