Thermocycler and sample port

• US 9,352,322 B2
• Filed: 01/30/2012
• Issued: 05/31/2016
• Est. Priority Date: 02/02/2006
• Status: Expired due to Fees

• Current Assignee: QIAGEN Instruments AG (Qiagen NV)
• Original Assignee: QIAGEN Instruments AG (Qiagen NV)
• Inventors: Stanley, Keith, Corbett, John
• Primary Examiner(s): MERKLING, SALLY ANNE
• Application Number:
  US13/361,315
• Publication Number:
  US 20120190074A1
• Time in Patent Office: 1,583 Days
• US Class Current: 1/1
• 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 continuous flow apparatus adapted to carry out nucleic acid amplification reactions, said continuous flow apparatus comprising:

(i) a continuous flow tube comprising a first inlet portion and an outlet; and

(ii) a sample port for introducing a volume of a liquid sample into a fluid carrier stream flowing through said continuous flow tube, said sample port comprising a reservoir for continuously supplying said first inlet portion of said continuous flow tube with said fluid carrier, said reservoir adapted to maintain a substantially constant level of fluid carrier above said first inlet portion and fluidly engaged with said first inlet portion,

wherein the first inlet portion is arranged with a longitudinal axis thereof extending along a direction of gravity, and

wherein said first inlet portion of said continuous flow tube is configured so as to be submerged within the fluid carrier contained in said reservoir and is vertically configured with respect to said reservoir to receive said liquid sample from above, 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 said liquid sample introduced into said carrier.

2. A continuous flow apparatus according to claim 1, which is a thermocycling apparatus.

3. A continuous flow apparatus according to claim 1, wherein the liquid sample is an aqueous sample.

4. A continuous flow apparatus according to claim 1, wherein the carrier fluid is a hydrophobic liquid.

5. A continuous flow apparatus according to claim 4, wherein the hydrophobic liquid is an oil.

6. A continuous flow apparatus according to claim 5, wherein the oil is a silicon oil or a silicon-based oil.

7. A continuous flow apparatus according to claim 1, wherein said reservoir comprises a bottom frusto-conical portion, wherein said first inlet portion extends through a base of said frusto-conical portion of said reservoir.

8. A continuous flow apparatus according to claim 1, further comprising a pump to supply said reservoir with said fluid carrier.

9. A continuous flow apparatus according to claim 1, further comprising a weir for maintaining the level of said fluid carrier in said reservoir.

10. A continuous flow apparatus according to claim 1, further comprising a vacuum arrangement engaged with the outlet of said continuous flow tube to apply a suction force to said outlet to draw said liquid sample through said continuous flow tube.

11. A continuous flow apparatus according to claim 1,

wherein the sample port further comprises a second inlet through which fluid carrier is introduced, and

wherein the reservoir has a top portion and a bottom portion, and said first inlet portion is arranged closer to the bottom portion than said second inlet portion.

12. A continuous flow apparatus according to claim 11, wherein the top portion of the reservoir is open to the atmosphere.

13. A continuous flow apparatus according to claim 1, wherein the reservoir has walls defining an interior volume containing the fluid carrier, and said first inlet portion extends through one or more walls of the reservoir into the volume of said reservoir.

14. A continuous flow apparatus according to claim 6, wherein the silicon oil has a viscosity of between about 5 to 50 centistokes.