Thermal cycler including a temperature gradient block

• US 6,962,821 B2
• Filed: 12/05/2002
• Issued: 11/08/2005
• Est. Priority Date: 10/20/1993
• Status: Expired due to Fees

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Stratagene California (Agilent Technologies, Inc.)
• Inventors: Danssaert, John Lewis, Shoemaker, Daniel Davis, Shopes, Robert James
• Primary Examiner(s): FREDMAN, JEFFREY NORMAN
• Application Number:
  US10/309,814
• Publication Number:
  US 20030157563A1
• Time in Patent Office: 1,069 Days
• Field of Search: 435/4, 435/288.4, 436/501, 422/99, 422/63, 422/67
• US Class Current: 436/501
• CPC Class Codes: B01L 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1838 using fluid heat transfer medium
• CPC Class Codes: B01L 2300/1844 using fans
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: B01L 7/5255 by moving sample containers
• CPC Class Codes: B01L 7/54 using spatial temperature gradients
• CPC Class Codes: G01N 2035/042 moved independently, e.g. by fork manipulator
• CPC Class Codes: G01N 35/0099 comprising robots or similar manipulators robots per se B25J
• CPC Class Codes: G01N 35/028 having reaction cells in the form of microtitration plates
• CPC Class Codes: Y10S 435/809 Incubators or racks or holders for culture plates or containers
• CPC Class Codes: Y10T 436/25 including sample preparation

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.

Claims

1. A method for simultaneously reacting a plurality of reaction mixtures in an apparatus including a temperature gradient block comprising the steps of:

placing reaction mixtures in a plurality of reaction wells in said gradient block, said gradient block having a top portion, first and second opposing portions, and a bottom portion, said plurality of reaction mixture wells being formed in said block between said opposing portions,

heating said gradient block using a resistive heater, and

generating a temperature gradient across said gradient block and between said opposing portions.

2. A method according to claim 1 wherein said step of generating a temperature gradient comprises the steps of heating said first opposing portion of said gradient block, and cooling said second opposing portion of said gradient block.

3. A method according to claim 1 including the further step of controlling said temperature gradient using controlling means.

4. A method according to claim 3 wherein said controlling step comprises the steps of collecting and storing temperature set point and actual temperature data from said wells, and transmitting said information to a microprocessor.

5. A method according to claim 1 wherein said apparatus further comprises at least one additional heat conducting block having a top portion, first and second opposing portions, and a bottom portion, and a plurality of reaction mixture wells formed in said additional block between said opposing portions, the method further comprising the step of moving said reaction mixtures between said gradient block and said additional block or blocks.

6. A method for automated temperature cycling of a reaction mixture using a thermal cycling apparatus comprising at least one heat conducting block, said block having a plurality of sample wells spaced between first and second opposing portions and in an upper surface thereof, the method comprising placing reaction mixtures in said wells, and heating said gradient block using a resistive heater to generate a temperature gradient across said block and between said opposing portions by heating said first opposing portion and cooling said second opposing portion.