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Rapid thermocycler system for rapid amplification of nucleic acids and related methods

  • US 9,737,891 B2
  • Filed: 05/31/2012
  • Issued: 08/22/2017
  • Est. Priority Date: 06/01/2011
  • Status: Active Grant
First Claim
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1. A method for operating a thermocycler instrument for amplification by a polymerase chain reaction, comprising the steps of:

  • a) operating the thermocycler instrument including;

    (i) a sample holder which is a block with a plurality of bores therein, wherein each bore is adapted to complementarily receive a sample tube,(ii) a pair of thermoelectric devices opposed and spaced apart having the sample holder disposed between, each of the thermoelectric devices having at least one heat exchanger,(iii) at least one first temperature sensor associated with the sample holder, and(iv) at least one second temperature sensor associated with the at least one beat exchanger,for causing at least one sample in the sample tube contained in the sample holder to undergo polymerase chain reaction amplification by repealed cycling between at least a denaturation heating stage and an annealing cooling stage;

    b) monitoring a first temperature corresponding with a temperature of the sample holder using the at least one first temperature sensor, and a second temperature corresponding with a temperature of the at least one heat exchanger and external of the sample holder using the at least one second temperature sensor, and wherein substantially real time temperature condition information obtained from the at least one first temperature sensor and the at least one second temperature sensor is employed and processed for delivering suitable power;

    c) dynamically controlling the power that is delivered to the pair of thermoelectric devices of the thermocycler instrument based upon the first temperature and the second temperature, wherein the dynamically controlling step (c) includes the steps of;

    (i) receiving at least one first setpoint temperature predetermined by a user, the at least one first setpoint temperature being a maximum temperature to which the at least one sample is to be heated for polymerase chain reaction denaturation of the at least one sample;

    (ii) receiving at least one second setpoint temperature predetermined by the user, the at least one second setpoint temperature being a minimum temperature to which the at least one sample is to be cooled for annealing of the at least one sample;

    (iii) receiving at least one first hold time corresponding with an amount of time predetermined by the user during which the temperature of the sample holder is maintained generally at the at least one first setpoint temperature;

    (iv) receiving at least one second hold time corresponding with an amount of time predetermined by the user during which the temperature of the sample holder is maintained generally at the at least one second setpoint temperature;

    (v) causing at least one of the thermoelectric devices to heat the sample holder;

    (vi) receiving a first sensor signal from the at least one first temperature sensor corresponding with the temperature of the sample holder;

    (vii) receiving a second sensor signal from the at least one second temperature sensor corresponding with the temperature of the at least one heat exchanger and is located externally of the sample holder;

    (viii) determining a value of any first temperature offset amount (TOFFSET1) based upon the temperature reading from the at least one first and second temperature sensors;

    (ix) causing heating of the sample holder until the sample holder reaches a first offset temperature that is below the first setpoint temperature by the first temperature offset amount;

    (x) at the time the first offset temperature is reached, causing a cooling pulsation of at least one of the thermoelectric devices during the heating stage for a sufficient amount of time so that the temperature arrives within about 1°

    C. of the first setpoint temperature and further heating of the sample holder is interrupted;

    (xi) causing the temperature to be maintained within about 1°

    C. of the first setpoint temperature for the first hold time;

    (xii) causing at least one of the thermoelectric devices to cool the sample holder until the temperature of the sample holder reaches a second offset temperature that is above the second setpoint temperature by a second temperature offset (TOFFSET2) amount;

    (xiii) receiving a temperature reading from the at least one first temperature sensor corresponding with the temperature of the sample holder;

    (xiv) receiving a temperature reading from the at least one second temperature sensor corresponding with the temperature of the at least one heat exchanger and is located remotely from the sample holder;

    (xv) determining a value of any second temperature offset amount (TOFFSET2) based upon the temperature reading from the at least one first and second temperature sensors;

    (xvi) causing cooling of the sample holder until the sample holder reaches the second offset temperature that is above the second setpoint temperature by the second temperature offset amount;

    (xvii) at the time when the second offset temperature is reached, causing a heating pulsation of at least one of the thermoelectric devices during the cooling stage for a sufficient amount of time so that the temperature arrives within about 1°

    C. of the second setpoint temperature and further cooling of the sample holder is interrupted;

    (xviii) causing the temperature to be maintained within about 1°

    C. of the second setpoint temperature for the second hold time; and

    (xix) repeating at least steps (v)-(xviii) for a predetermined number of cycles until the amplification desired by the user is achieved.

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