THERMOCYCLER AND SAMPLE PORT

• US 20090220966A1
• Filed: 02/02/2007
• Published: 09/03/2009
• Est. Priority Date: 02/02/2006
• Status: Active Grant

• Current Assignee: QIAGEN Instruments AG (Qiagen NV)
• Original Assignee: Corbett Life Science (Qiagen NV)
• Inventors: Stanley, Keith, Corbett, John
• Granted Patent: US 8,124,413 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/0838 Capillaries
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: B01L 7/54 using spatial temperature gradients
• CPC Class Codes: Y10T 137/0318 Processes
• CPC Class Codes: Y10T 436/118339 with formation of a segmented stream
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

The invention relates to continuous flow systems, in particular thermocyclers for the automated and continuous cycling of fluid between a plurality of temperature zones in the amplification of nucleic acids. The invention also relates to an improved sample port for introducing a volume of a liquid sample into a continuous flow system.

Claims

1. A thermocycler comprising a plurality of nested heat exchangers for independently maintaining a predetermined temperature thereby to define a corresponding plurality of temperature zones;

said heat exchangers adapted to receive a reaction tube such that said reaction tube is in heat transfer communication with said heat exchangers, whereby a fluid passing through a reaction tube engaged with said heat exchangers passes cyclically through the temperature zones.

2. A method of amplifying a nucleic acid in a PCR or LCR format comprising using the thermocycler according to claim 1.

3. A method of performing a nucleic acid melt detection assay comprising using the thermocycler according to claim 1.

4. A nucleic acid prepared using the thermocycler according to claim 1.

15. A thermocycler according to claim 1 further comprising a scanning detector for monitoring the course of a reaction occurring in said reaction tube.

16. A thermocycler according to claim 15 wherein said reaction is measured by fluorescence.

17. A thermocycler according to claim 15 wherein said scanning detector comprises a rotatable unit axially mounted either above or below or within an inner heat exchanger ring for directly measuring said reaction occurring in said reaction tube.

18. A thermocycler according to claim 17 wherein said reaction tube is directly measured in a gap between said inner and outer heat exchangers and/or through a viewing slot.

19. A thermocycler according to claim 15 wherein a plurality of detectors are mounted above the gap between said nested heat exchangers for monitoring the course of a reaction occurring in said reaction tube, wherein an equal number of detectors are provided for the number of exposed reaction tubes.

5. A sample port for introducing a volume of a liquid sample into a fluid carrier stream flowing through a continuous flow tube having an outlet and a common inlet into which said carrier stream and said liquid sample are both introduced, said port comprising:

a reservoir for continuously supplying said inlet with said fluid carrier, said reservoir adapted to maintain a substantially constant level of fluid carrier above said inlet and fluidly engageable with said inlet of said continuous flow tube such that, in use, said fluid carrier stream and said liquid sample are drawn through said continuous flow tube when said reservoir is at substantially atmospheric pressure and when said fluid carrier is chosen such that its properties are sufficient to maintain the physical shape of the liquid sample introduced into said carrier.

6. A sample port according to claim 5, wherein the liquid sample is an aqueous sample.

7. A sample port according to claim 5, wherein the carrier fluid is a hydrophobic liquid.

8. A sample port according to claim 7, wherein the hydrophobic liquid is an oil.

9. A sample port according to claim 8, wherein the oil is a silicon oil or a silicon-based oil.

10. A method for introducing a volume of a liquid sample into a fluid carrier stream flowing through a continuous flow tube having an outlet and a common inlet into which said carrier stream and said liquid sample are both introduced, said method comprising the steps of:

providing a sample port according to claim 5;

fluidly engaging said inlet of said continuous flow tube with said reservoir;

introducing said fluid carrier into said reservoir and introducing said liquid sample into said fluid carrier, said fluid carrier chosen such that its properties are sufficient to maintain the physical shape of the liquid sample and such that said fluid carrier stream and said liquid sample are drawn through said continuous flow tube when said reservoir is at substantially atmospheric pressure.

11. A method for introducing a volume of a liquid sample into a fluid carrier stream flowing through a continuous flow tube having an outlet and a common inlet into which said carrier stream and said liquid sample are both introduced, said method comprising the steps of:

providing a port according to claim 5;

fluidly engaging said inlet of said continuous flow tube with said reservoir;

introducing said fluid carrier into said reservoir;

immersing a liquid sample dispenser into said fluid carrier contained in said reservoir;

dispensing said liquid sample adjacent said inlet and optionally maneuvering said dispensed liquid sample with said liquid sample dispenser such that said dispensed liquid sample is introduced into said inlet and drawn through said continuous flow tube, said fluid carrier chosen such that its properties are sufficient to maintain the physical shape of the liquid sample and such that said fluid carrier stream and said liquid sample are drawn through said continuous flow tube when said reservoir is at substantially atmospheric pressure.

12. A continuous flow apparatus comprising the sample port according to claim 5.

13. A continuous flow apparatus according to claim 12, which is a thermocycling apparatus for performing nucleic acid amplification reactions.

14. A method of amplifying a nucleic acid in a PCR or LCR format comprising using the continuous flow apparatus according to claim 13.