System and method for automated nucleic acid amplification

• US 9,808,806 B2
• Filed: 08/25/2014
• Issued: 11/07/2017
• Est. Priority Date: 08/26/2013
• Status: Active Grant

• Current Assignee: 454 Life Sciences Corporation (Roche Holding AG)
• Original Assignee: 454 Life Sciences Corporation (Roche Holding AG)
• Inventors: Tajima, Hideji, Ueda, Tetsuya, Kobayashi, Shuichi, Kane, Nathan Robert
• Primary Examiner(s): Priest, Aaron
• Application Number:
  US14/467,321
• Publication Number:
  US 20150056663A1
• Time in Patent Office: 1,170 Days
• Field of Search: None
• CPC Class Codes: B01F 23/41 Emulsifying
• CPC Class Codes: B01F 27/0721 parallel with respect to the rotating axis
• CPC Class Codes: B01F 27/0724 directly mounted on the rotating axis
• CPC Class Codes: B01F 27/213 characterised by the connection with the drive
• CPC Class Codes: B01F 31/445 performing an oscillatory movement about an axis
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/0809 rectangular shaped
• CPC Class Codes: B01L 2300/0858 Side walls
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 3/0293 for liquids
• CPC Class Codes: B01L 3/508 rigid containers not provided for above
• 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: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 2035/00534 Mixing by a special element, e.g. stirrer
• CPC Class Codes: G01N 2035/00891 Displaying information to the operator

Abstract

An embodiment of a device for automatically executing a process of generating an emulsion containing nucleic acids, amplifying the nucleic acids in the emulsion, breaking the emulsion, and separating and purifying said amplified nucleic acids, is described that comprises an emulsion generation unit for sealing beads to which nucleic acids are bound in a water-in-oil type emulsion; a nucleic acid amplification unit provided with a reaction vessel for amplifying said nucleic acids and a heating and cooling part for heating and cooling the reaction vessel; an emulsion breaking unit for breaking the emulsion after nucleic acid amplification; and a nucleic acid purification unit for recovering said amplified nucleic acids from said emulsion breaking unit.

Claims

1. A device for automatically executing a process of generating an emulsion containing nucleic acids, amplifying the nucleic acids in the emulsion, breaking the emulsion, and separating and purifying said amplified nucleic acids, said device comprising:

an emulsion generation unit for sealing beads to which nucleic acids are bound in a water-in-oil type emulsion, wherein the emulsion generation unit comprises a container;

a nucleic acid amplification unit provided with a reaction vessel for amplifying said nucleic acids and a heating and cooling part for heating and cooling the reaction vessel;

an emulsion breaking unit for breaking the emulsion after nucleic acid amplification, wherein the emulsion breaking unit comprises a container and is connected to the reaction vessel by a channel; and

a nucleic acid purification unit for recovering said amplified nucleic acids from said emulsion breaking unit, wherein the nucleic acid purification unit comprises multiple tubes,

wherein a head drop is disposed between said nucleic acid amplification unit and said emulsion generation unit and/or between said emulsion breaking unit and said nucleic acid amplification unit via a plurality of liquid transfer channels for liquid transfer, and

wherein.

said nucleic acid amplification unit is disposed below said emulsion generation unit, and said emulsion breaking unit is disposed below said nucleic acid amplification unit.

2. The device of claim 1, wherein

a channel opening and closing device is provided on said liquid transfer channels.

3. The device of claim 1, wherein

said liquid transfer channels are formed from soft tubes, hoses, or films.

4. The device of claim 1, wherein

said liquid transfer channels are molded integrally with or attached to at least one of said units.

5. The device of claim 1, wherein

said reaction vessel comprises a bag.

6. The device of claim 1, wherein

said nucleic acid amplification unit is covered by a heat-insulating material.

7. The device of claim 1 further comprising

a fluid volume sensor for detecting the fluid volume inside at least one of said units.

8. The device of claim 1, further comprising

a frame for holding at least one of said emulsion generation unit, said nucleic acid amplification unit, and said emulsion breaking unit.

9. The device of claim 1, further comprising

an automatic processing mechanism for performing prescribed processing selected from the group consisting of dispensing, stirring, and suction processing of the emulsion or solution inside at least one of said emulsion generation unit, said nucleic acid amplification unit, said emulsion breaking unit, and said nucleic acid purification unit, wherein said automatic processing mechanism comprises a dispensing burette, a magnet for adsorbing a plurality of magnetic beads, a stirring instrument for stirring the contents of at least one of said units, a nozzle unit, or combinations thereof.

10. The device of claim 9, further comprising

a moving mechanism for moving said automatic processing mechanism three-dimensionally.

11. The device of claim 10, wherein

said moving mechanism moves said automatic processing mechanism to at least one of the positions of said emulsion generation unit, said nucleic acid amplification unit, said emulsion breaking unit, and said nucleic acid purification unit.

12. The device of claim 11, further comprising

an operating stage on an upper surface of the device of claim 1;

wherein said moving mechanism moves said automatic processing mechanism three-dimensionally on said operating stage.

13. The device of claim 10, wherein

said moving mechanism comprises a first direction driving part for driving said automatic processing mechanism in a first direction, a second direction driving part for driving said automatic processing mechanism in a second direction differing from said first direction, and a third direction driving part for driving said automatic processing mechanism in a third direction differing from said first and second directions.

14. The device of claim 13, wherein

said first direction of said first direction driving part is a direction inclined with respect to a horizontal plane.

15. A method for generating an emulsion containing nucleic acids, amplifying the nucleic acids in the emulsion, breaking the emulsion, and separating and purifying said amplified nucleic acids using the device of claim 1.