Apparatus, system, and method using immiscible-fluid-discrete-volumes

• US 9,194,772 B2
• Filed: 09/10/2009
• Issued: 11/24/2015
• Est. Priority Date: 08/22/2005
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Lee, Linda, Woo, Sam, Ma, Congcong, Reel, Richard, Oldham, Mark, Cox, David, Schroeder, Benjamin, Sorenson, Jon, Wiyatno, Willy
• Primary Examiner(s): Wilder, Cynthia B
• Application Number:
  US12/557,488
• Publication Number:
  US 20100209916A1
• Time in Patent Office: 2,266 Days
• Field of Search: 435/91.2, 435/6.11, 435/287.2
• US Class Current: 1/1
• CPC Class Codes: B01J 2219/00353 Pumps
• CPC Class Codes: B01J 2219/0036 Nozzles
• CPC Class Codes: B01J 2219/00364 Pipettes
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00619 using hydrophilic or hydrophobic regions
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00653 the compounds being bound to electrodes embedded in or on the solid supports
• CPC Class Codes: B01J 2219/00657 One-dimensional arrays
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/0293 for liquids
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502784 : specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• 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 1/6806 : Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6869 : Methods for sequencing
• CPC Class Codes: C12Q 1/6874 : involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2535/101 : Sanger sequencing method, i.e. oligonucleotide sequencing using primer elongation and dideoxynucleotides as chain terminators
• CPC Class Codes: F15C 5/00 : Manufacture of fluid circuit elements; Manufacture of assemblages of such elements integrated circuits
• CPC Class Codes: F16K 2099/0084 : Chemistry or biology, e.g. "lab-on-a-chip" technology
• CPC Class Codes: F16K 99/0001 : Microvalves microdevices B81B1/00; manufacture or treatment of devices or systems in or on a substrate B81C1/00; microfluidic structures B01L3/5027; micropumps F04B19/006
• CPC Class Codes: F16K 99/0011 : Gate valves or sliding valves
• CPC Class Codes: F16K 99/0013 : Rotary valves
• CPC Class Codes: G01N 1/14 : Suction devices, e.g. pumps; Ejector devices
• CPC Class Codes: G01N 27/44743 : Introducing samples
• CPC Class Codes: G01N 27/44769 : Continuous electrophoresis, i.e. the sample being continuously introduced, e.g. free flow electrophoresis [FFE]
• CPC Class Codes: G01N 35/08 : using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
• CPC Class Codes: Y10T 137/0318 : Processes
• CPC Class Codes: Y10T 137/4259 : With separate material addition
• CPC Class Codes: Y10T 137/85978 : With pump
• CPC Class Codes: Y10T 137/85986 : Pumped fluid control
• CPC Class Codes: Y10T 137/86863 : Rotary valve unit
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

Various embodiments of the teachings relate to a system or method for sample preparation or analysis in biochemical or molecular biology procedures. The sample preparation can involve small volume processed in discrete portions or segments or slugs, herein referred to as discrete volumes. A molecular biology procedure can be nucleic acid analysis. Nucleic acid analysis can be an integrated DNA amplification/DNA sequencing procedure.

Claims

1. A system comprising:

at least one conduit;

an aqueous sample fluid introduction unit in fluid communication with the at least one conduit;

a spacing fluid introduction unit in fluid communication with the at least one conduit, the aqueous sample fluid introduction unit and the spacing fluid introduction unit comprising separate units each in fluid communication with the at least one conduit;

a control unit configured to control a flow an aqueous sample fluid and a spacing fluid from the aqueous sample fluid introduction unit and the spacing fluid introduction unit, respectively, to form discrete volumes of the aqueous sample fluid within the spacing fluid in the at least one conduit, wherein the aqueous sample fluid contains at least one nucleic acid fragment, and the sample fluid introduction unit is further configured to inject volumes of aqueous sample fluid such that at least 1% of the discrete volumes contain a single nucleic acid fragment; and

a detector configured to detect the at least one nucleic acid fragment within the discrete volumes of aqueous sample flowing through the at least one conduit.

2. The system of claim 1, wherein an inner surface of the at least one conduit comprises an attached linker moiety.

3. The system of claim 1, further comprising an aqueous sample fluid source in fluid communication with the aqueous sample fluid introduction unit, and a spacing fluid source in fluid communication with the spacing fluid introduction unit.

4. The system of claim 1, wherein the detector is configured to detect a component of the aqueous sample fluid source, or a product thereof, while in the conduit.

5. The system of claim 4, wherein the detector comprises at least one of a spectrophotometer, a refractive index detector, and a fluorometer.

6. The system of claim 1, further comprising a thermal-cycling device adapted to thermally cycle an aqueous sample fluid in the at least one conduit.

7. The system of claim 1, wherein the conduit comprises a conduit having an inner diameter and the inner diameter is about 300 microns or less.

8. The system of claim 7, wherein the inner diameter is about 50 microns or less.