Apparatus and method for performing automated amplification of nucleic acid sequences and assays using heating and cooling steps

Claim 1

An apparatus for performing temperature cycling of a reaction mix comprising: a heat-conducting container for holding a reaction mixture; means for heating and cooling said container to or at any of a plurality of user-defined temperatures and having a control input for receiving a control signal controlling whether said container is heated or cooled; and a computer means, coupled to said control input of said means, for receiving and storing checkpoint data from the user defining the plurality of temperatures and the times at which said temperatures are to be attained thereby defining a temperature profile, and for, upon receipt of a command from the user, accessing said checkpoint data and generating control signals therefrom at the control input of said means for heating and cooling to cause the user-defined temperature profile to be achieved at said container; and wherein said means for heating and cooling is an aluminum plate having fluid flow channels formed therein which are in fluid communication with pumps which circulate fluid stored in fluid reservoirs having heating and cooling elements therein to keep the fluid at a constant, user-definable temperature.

Claim 2

An apparatus for performing temperature cycling of a reaction mix comprising: a heat-conducting container for holding a reaction mixture; means for heating and cooling said container to or at any of a plurality of user-defined temperatures and having a control input for receiving a control signal controlling whether said container is heated or cooled; and a computer means, coupled to said control input of said means, for receiving and storing checkpoint data from the user defining the plurality of temperatures and the times at which said temperatures are to be attained thereby defining a temperature profile, and for, upon receipt of a command from the user, accessing said checkpoint data and generating control signals therefrom at the control input of said means for heating and cooling to cause the user-defined temperature profile to be achieved at said container; and wherein said means for heating and cooling is an aluminum plate having fluid flow channels formed therein which are in fluid communication with pumps which circulate fluid stored in fluid reservoirs having heating and cooling elements therein to keep the fluid at a constant, user-definable temperature.

Claim 3

An apparatus for performing temperature cycling of a reaction mix comprising: a heat-conducting container for holding a reaction mixture; means for heating and cooling said container to or at any of a plurality of user-defined temperatures and having a control input for receiving a control signal controlling whether said container is heated or cooled; and a computer means, coupled to said control input of said means, for receiving and storing checkpoint data from the user defining the plurality of temperatures and the times at which said temperatures are to be attained thereby defining a temperature profile, and for, upon receipt of a command from the user, accessing said checkpoint data and generating control signals therefrom at the control input of said means for heating and cooling to cause the user-defined temperature profile to be achieved at said container; and wherein said computer means includes means for receiving and storing in a link data field in a database associated with each said temperature profile stored by said computer means link data entered by the user for every set of checkpoints defining a temperature profile, and for receiving and storing a plurality of sets of checkpoints input by the user to define a plurality of temperature profiles, each of which has its own link data item, and wherein said computer means also includes means to run any particular temperature profile identified, if any is identified, in the link data field of the temperature profile just run and to continue this process of running the temperature profiles identified in the link data fields associated with each temperature profile run until no more temperature profiles are identified.

Claim 4

An apparatus for performing temperature cycling of a reaction mix comprising: a heat-conducting container for holding a reaction mixture; means for heating and cooling said container to or at any of a plurality of user-defined temperatures and having a control input for receiving a control signal controlling whether said container is heated or cooled; and a computer means, coupled to said control input of said means, for receiving and storing checkpoint data from the user defining the plurality of temperatures and the times at which said temperatures are to be attained thereby defining a temperature profile, and for, upon receipt of a command from the user, accessing said checkpoint data and generating control signals therefrom at the control input of said means for heating and cooling to cause the user-defined temperature profile to be achieved at said container; and wherein said computer means includes means for receiving and storing in a link data field in a database associated with each said temperature profile stored by said computer means link data entered by the user for every set of checkpoints defining a temperature profile, and for receiving and storing a plurality of sets of checkpoints input by the user to define a plurality of temperature profiles, each of which has its own link data item, and wherein said computer means also includes means to run any particular temperature profile identified, if any is identified, in the link data field of the temperature profile just run and to continue this process of running the temperature profiles identified in the link data fields associated with each temperature profile run until no more temperature profiles are identified.

Claim 5

An apparatus for performing temperature cycling of a reaction mix comprising: a heat-conducting container for holding a reaction mixture; means for heating and cooling said container to or at any of a plurality of user-defined temperatures and having a control input for receiving a control signal controlling whether said container is heated or cooled; and a computer means, coupled to said control input of said means, for receiving and storing checkpoint data from the user defining the plurality of temperatures and the times at which said temperatures are to be attained thereby defining a temperature profile, and for, upon receipt of a command from the user, accessing said checkpoint data and generating control signals therefrom at the control input of said means for heating and cooling to cause the user-defined temperature profile to be achieved at said container; and wherein said computer means includes means for receiving and storing in a link data field in a database associated with each said temperature profile stored by said computer means link data entered by the user for every set of checkpoints defining a temperature profile, and for receiving and storing a plurality of sets of checkpoints input by the user to define a plurality of temperature profiles, each of which has its own link data item, and wherein said computer means also includes means to run any particular temperature profile identified, if any is identified, in the link data field of the temperature profile just run and to continue this process of running the temperature profiles identified in the link data fields associated with each temperature profile run until no more temperature profiles are identified; and further comprising means in said computer means for receiving and storing data for a number of cycles data field from the user for each set of checkpoints entered by the user to define a temperature profile and for running each temperature profile the number of times identified in said number of cycles data field before checking said link field of the temperature profile for the identification of the temperature profile to be run next.

Claim 6

An apparatus for performing temperature cycling of a reaction mix comprising: a heat-conducting container for holding a reaction mixture; means for heating and cooling said container to or at any of a plurality of user-defined temperatures and having a control input for receiving a control signal controlling whether said container is heated or cooled; and a computer means, coupled to said control input of said means, for receiving and storing checkpoint data from the user defining the plurality of temperatures and the times at which said temperatures are to be attained thereby defining a temperature profile, and for, upon receipt of a command from the user, accessing said checkpoint data and generating control signals therefrom at the control input of said means for heating and cooling to cause the user-defined temperature profile to be achieved at said container; and wherein said computer means includes means for receiving and storing in a link data field in a database associated with each said temperature profile stored by said computer means link data entered by the user for every set of checkpoints defining a temperature profile, and for receiving and storing a plurality of sets of checkpoints input by the user to define a plurality of temperature profiles, each of which has its own link data item, and wherein said computer means also includes means to run any particular temperature profile identified, if any is identified, in the link data field of the temperature profile just run and to continue this process of running the temperature profiles identified in the link data fields associated with each temperature profile run until no more temperature profiles are identified; and further comprising means in said computer means for receiving and storing data for a number of cycles data field from the user for each set of checkpoints entered by the user to define a temperature profile and for running each temperature profile the number of times identified in said number of cycles data field before checking said link field of the temperature profile for the identification of the temperature profile to be run next.

Claim 7

An apparatus for performing automated amplification of at least one specific nucleic acid sequence comprising: a first means for holding a reaction mixture comprising said nucleic acid sequence(s) to be amplified, four different nucleotide triphosphates, a thermostable enzyme, and one oligonucleotide primer for each different specific sequence being amplified, wherein each primer is selected to be substantially complementary to different strands of each specific sequence, such that the extension product synthesized from one primer, when it is separated from its complement, can serve as a template for synthesis of the extension product of the other primer, said holding being carried out at any selected temperature or plurality of temperatures; and a second means for automatically performing a predetermined sequence of steps including causing said first means to heat its contents for a first period and to cool its contents for a second period; and wherein said second means includes means for allowing the user to enter data which control certain process parameters that characterize predetermined steps in said sequence of steps; and wherein said first means holds said liquid stored therein at either of two temperatures and wherein said second second means causes said first means to hold its contents at a first temperature for a user-defined interval followed by a chill-down period and a low-temperature incubation at said second temperature having a user-defined duration; and wherein said first means is a reaction chamber thermally coupled to an aluminum plate having fluid flow channels formed therein which are in fluid communication with pumps which circulate fluid stored in fluid reservoirs having heating and cooling elements therein to keep the fluid at a constant, user-definable temperature.

Claim 8

An apparatus for performing automated amplification of at least one specific nucleic acid sequence comprising: a first means for holding a reaction mixture comprising said nucleic acid sequence(s) to be amplified, four different nucleotide triphosphates, a thermostable enzyme, and one oligonucleotide primer for each different specific sequence being amplified, wherein each primer is selected to be substantially complementary to different strands of each specific sequence, such that the extension product synthesized from one primer, when it is separated from its complement, can serve as a template for synthesis of the extension product of the other primer, said holding being carried out at any selected temperature or plurality of temperatures; and a second means for automatically performing a predetermined sequence of steps including causing said first means to heat its contents for a first period and to cool its contents for a second period; and wherein said second means includes means for allowing the user to enter data which control certain process parameters that characterize predetermined steps in said sequence of steps; and wherein said first means holds said liquid stored therein at either of two temperatures and wherein said second second means causes said first means to hold its contents at a first temperature for a user-defined interval followed by a chill-down period and a low-temperature incubation at said second temperature having a user-defined duration; and wherein said first means is a reaction chamber thermally coupled to an aluminum plate having fluid flow channels formed therein which are in fluid communication with pumps which circulate fluid stored in fluid reservoirs having heating and cooling elements therein to keep the fluid at a constant, user-definable temperature.