Method for rapid thermal cycling of biological samples

• US 20040265892A1
• Filed: 07/13/2004
• Published: 12/30/2004
• Est. Priority Date: 06/04/1990
• Status: Active Grant

• Current Assignee: Carl T. Wittwer, David R. Hillyard
• Original Assignee: Carl T. Wittwer, David R. Hillyard
• Inventors: Hillyard, David R., Wittwer, Carl T.
• Granted Patent: US 7,238,321 B2
• Time in Patent Office: Days
• US Class Current: 435/6.12
• CPC Class Codes: B01D 15/34 Size selective separation, e.g. size exclusion chromatography, gel filtration, permeation
• CPC Class Codes: B01L 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2300/0838 Capillaries
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1844 using fans
• CPC Class Codes: B01L 2300/1872 Infrared light
• CPC Class Codes: B01L 2300/1883 using thermal insulation
• 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 9/065 specially adapted for capillary tubes
• CPC Class Codes: G01N 27/447 using electrophoresis
• CPC Class Codes: G01N 30/02 Column chromatography
• CPC Class Codes: G01N 30/06 Preparation

Abstract

A thermal cycling method and device is disclosed. The device comprises a sample chamber whose temperature can be rapidly and accurately modulated over a range of temperatures needed to carry out a number of biological procedures, such a DNA polymerase chain reaction. Biological samples are placed in glass micro capillary tubes and then located inside the sample chamber. A programmable controller regulates the temperature of the sample inside the sample chamber. Once a heating cycle is completed, the controller opens a door to the chamber for venting hot air out and cool ambient air is moved in. Temperature versus time profiles corresponding to optimum denaturation, annealing and elongation temperatures for amplification of DNA are achieved by the present invention.

Claims

1. A thermal cycler for amplifying a nucleic acid comprising:

a housing comprising a chamber;

a port in said housing for receiving at least one PCR sample holder in said chamber;

a heating assembly for heating a sample comprising the nucleic acid, said heating assembly in thermal communication with said PCR sample holder when said sample holder is present in said chamber;

a cooling assembly for cooling the sample, said cooling assembly thermal communication with said PCR sample holder when said PCR sample holder is present in said chamber; and

a controller for said heating and said cooling assemblies wherein said controller is capable of cycling the sample through a temperature cycle within a time range of about 30-60 seconds corresponding to the denaturation, annealing and elongation steps of a polymerase chain reaction and wherein said PCR sample holder has a thermal mass which provides for completing said cycle within the time range of about 30-60 seconds.

2. The thermal cycler of claim 1 wherein said heating assembly comprises a resistance heater.

3. The thermal cycler of claim 2 wherein said resistance heater comprises a heating coil.

4. The thermal cycler of claim 1 wherein said heating assembly is in fluid communication with said PCR sample holder when said PCR sample holder is present in said chamber.

5. The thermal cycler of claim 4 wherein said heating assembly comprises a blower.

6. The thermal cycler of claim 1 wherein said cooling assembly is in fluid communication with said PCR sample holder when said PCR sample holder is present in said chamber.

7. The thermal cycler of claim 6 wherein said cooling assembly comprises a blower.

8. The thermal cycler of claim 6 wherein said chamber comprises a fluid inlet and a fluid outlet to provide for the entry of ambient air and exhaust of heated air to cool said PCR sample holder when said PCR sample holder is present in said chamber.

9. The thermal cycler of claim 1 wherein said controller is capable of lowering the temperature of said PCR sample holder at a rate of at least about 1.5 degrees Celsius per second.

10. The thermal cycler of claim 7 wherein the motor for said blower is external to said chamber

11. The thermal cycler of claim 1 in combination with said PCR sample holder.