Thermal cycler including a temperature gradient block
DCFirst Claim
1. A method for simultaneously reacting a plurality of reaction mixtures in an apparatus including a temperature gradient block and further including first and second uniform temperature blocks, said method comprising the steps of:
- placing reaction mixtures in a plurality of reaction wells in said gradient block, said gradient block having first and second opposing end portions, said plurality of reaction mixture wells being formed in said block between said opposing end portions;
generating a temperature gradient across said gradient block and between said opposing end portions; and
placing the reaction mixtures in a plurality of reaction wells in said first uniform temperature block; and
placing the reaction mixtures in a plurality of reaction wells in said second uniform temperature block; and
generating uniform temperatures in said uniform temperature blocks.
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Abstract
A method in which a temperature gradient is generated across a "gradient" block, and an apparatus comprising a block across which a temperature gradient can be generated. By setting up such a gradient, multiple reaction mixtures held in wells on the gradient block can be simultaneously run at temperatures which differ only slightly, thereby permitting an optimum temperature for the reaction to be quickly identified. In a preferred embodiment the gradient block is integrated into a thermal cycler used for nucleic acid amplification reactions.
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Citations
22 Claims
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1. A method for simultaneously reacting a plurality of reaction mixtures in an apparatus including a temperature gradient block and further including first and second uniform temperature blocks, said method comprising the steps of:
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placing reaction mixtures in a plurality of reaction wells in said gradient block, said gradient block having first and second opposing end portions, said plurality of reaction mixture wells being formed in said block between said opposing end portions; generating a temperature gradient across said gradient block and between said opposing end portions; and placing the reaction mixtures in a plurality of reaction wells in said first uniform temperature block; and placing the reaction mixtures in a plurality of reaction wells in said second uniform temperature block; and generating uniform temperatures in said uniform temperature blocks. - View Dependent Claims (2, 3, 4)
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5. An apparatus for generating a temperature gradient across a heat conducting block comprising:
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a reaction mixture holder including a heat conducting block having top and bottom portions, first and second opposing end portions, and a plurality of reaction mixture wells formed in said top portion; a first block heater positioned adjacent to said first opposing end portion; a block cooler positioned adjacent to said second opposing end portion; a second block heater positioned adjacent to said bottom portion; and means for independently controlling the first and second block heaters and the block cooler. - View Dependent Claims (6, 7, 8, 9)
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10. An apparatus for performing molecular biological reactions comprising:
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a temperature-gradient block, said temperature-gradient block having a plurality of reaction mixture wells spaced between first and second opposing end portions thereof; means for generating a temperature gradient between said first and second opposing end portions of the temperature-gradient block; first and second uniform-temperature blocks, each having a plurality of reaction mixture wells; and means for independently heating each uniform-temperature block. - View Dependent Claims (11, 12, 13, 14)
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15. A method for determining an optimum temperature for one step of a multistep biological reaction process comprising the steps of:
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generating a temperature gradient across a gradient-temperature block having a plurality of reaction wells, wherein the reaction wells are at different temperatures; placing reaction mixtures in the reaction wells in the gradient-temperature block to subject the reaction mixtures to different temperatures as one step of the multistep biological reaction process; generating a uniform temperature in a uniform-temperature block having a plurality of reaction wells; placing the reaction mixtures in the reaction wells in the uniform-temperature block as another step of the multistep biological reaction process; and analyzing the results of the multistep biological reaction process according to the different temperatures applied to the reaction mixtures in the gradient-temperature block to determine an optimum temperature for said one step. - View Dependent Claims (16, 17, 18, 19, 20, 21, 22)
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Specification