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.

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.

Claim 5

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.

Claim 6

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.

Claim 9

An apparatus for performing automated amplification of at least one specific nucleic acid sequence comprising: a reaction chamber; a solid state heat pump thermally coupled to said reaction chamber; an interface means having a heat control signal input for receiving a heat control signal and a cool control signal input for receiving a cool control signal, and having a power output coupled to said solid state heat pump for converting said heat and cool control signals into current flow through said solid state heat pump having the correct properties to cause either heating or cooling as indicated by said heat and cool control signals; and a computer means having heat and cool control signal outputs coupled to supply said heat and cool control signals to said interface means for receiving and storing one or more sets of checkpoints defining one or more temperature profiles, where each checkpoint is entered by the user and defines a specific temperature to be achieved or maintained at a specific time of the temperature profile with which it is associated, and for receiving and storing a number of cycles data field for each temperature profile defining how many times that temperature profile is to be run and for receiving link data from said user and storing it in a link data field indicating the next temperature profile to be run, said computer means also including means for, upon receiving a start command and a temperature profile identification from said user, running the identified temperature profile the number of times identified in said number of cycles data field by generating said heat and cool control signals in the proper sequence to cause the user-defined temperature profile to exist at said reaction chamber, and, after completing the desired number of cycles, for accessing the link data field of the temperature profile just completed and running the temperature profile identified, if any, in said link data field, and continuing this process of running linked temperature profiles until the link field of the temperature profile just completed does not identify another temperature profile to be run.

Claim 10

An apparatus for performing automated amplification of at least one specific nucleic acid sequence comprising: a reaction chamber; a solid state heat pump thermally coupled to said reaction chamber; an interface means having a heat control signal input for receiving a heat control signal and a cool control signal input for receiving a cool control signal, and having a power output coupled to said solid state heat pump for converting said heat and cool control signals into current flow through said solid state heat pump having the correct properties to cause either heating or cooling as indicated by said heat and cool control signals; and a computer means having heat and cool control signal outputs coupled to supply said heat and cool control signals to said interface means for receiving and storing one or more sets of checkpoints defining one or more temperature profiles, where each checkpoint is entered by the user and defines a specific temperature to be achieved or maintained at a specific time of the temperature profile with which it is associated, and for receiving and storing a number of cycles data field for each temperature profile defining how many times that temperature profile is to be run and for receiving link data from said user and storing it in a link data field indicating the next temperature profile to be run, said computer means also including means for, upon receiving a start command and a temperature profile identification from said user, running the identified temperature profile the number of times identified in said number of cycles data field by generating said heat and cool control signals in the proper sequence to cause the user-defined temperature profile to exist at said reaction chamber, and, after completing the desired number of cycles, for accessing the link data field of the temperature profile just completed and running the temperature profile identified, if any, in said link data field, and continuing this process of running linked temperature profiles until the link field of the temperature profile just completed does not identify another temperature profile to be run.

Claim 11

A method for amplifying at least one specific nucleic acid sequence comprising the steps of: using a computer-directed machine to heat to a predetermined. temperature for a predetermined time a sample of the 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 (hereafter the mixture), and using a computer-directed machine to heat the mixture to a user-defined temperature and to chill the mixture to a user-defined temperature.

Claim 12

A method for amplifying at least one specific nucleic acid sequence comprising the steps of: using a computer-directed machine to heat to a predetermined. temperature for a predetermined time a sample of the 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 (hereafter the mixture), and using a computer-directed machine to heat the mixture to a user-defined temperature and to chill the mixture to a user-defined temperature.

Claim 17

A method of amplifying at least one specific nucleic acid sequence comprising the steps of: a) using a computer-directed machine to issue a heat signal to a heating apparatus to cause a reaction chamber to be heated for a predetermined time to and/or at a predetermined temperature, wherein said reaction chamber contains a sample of the 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; b) using a computer-directed machine to issue a cool signal to a cooling apparatus to cause said reaction chamber to be cooled for a predetermined time to a predetermined temperature; and c) using a computer-directed machine to repeat the cycle consisting of steps a through c when the elapsed time for the active cooling signal equals a user-defined time if the number of cycles performed thus far is less than a user-defined number of cycles.

Claim 18

A method of amplifying at least one specific nucleic acid sequence comprising the steps of: a) using a computer-directed machine to issue a heat signal to a heating apparatus to cause a reaction chamber to be heated for a predetermined time to and/or at a predetermined temperature, wherein said reaction chamber contains a sample of the 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; b) using a computer-directed machine to issue a cool signal to a cooling apparatus to cause said reaction chamber to be cooled for a predetermined time to a predetermined temperature; and c) using a computer-directed machine to repeat the cycle consisting of steps a through c when the elapsed time for the active cooling signal equals a user-defined time if the number of cycles performed thus far is less than a user-defined number of cycles.