Methods for nucleic acid amplification on a printed circuit board

• US 8,048,628 B2
• Filed: 05/24/2007
• Issued: 11/01/2011
• Est. Priority Date: 09/24/2002
• Status: Expired due to Fees

• Current Assignee: Duke University
• Original Assignee: Duke University
• Inventors: Fair, Richard B., Pamula, Vamsee K., Pollack, Michael G.
• Primary Examiner(s): Horlick; Kenneth R.
• Application Number:
  US11/753,318
• Publication Number:
  US 20070217956A1
• Time in Patent Office: 1,622 Days
• Field of Search: None
• US Class Current: 435/6.12
• CPC Class Codes: B01F 33/3021 the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets
• CPC Class Codes: B01F 33/3031 using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids
• 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 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0663 Whole sensors
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• 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 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/089 Virtual walls for guiding liquids
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0424 Dielectrophoretic forces
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502792 for moving individual droplets on a plate, e.g. by locally altering surface tension
• 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: B81B 2201/058 Microfluidics not provided for in B81B2201/051 - B81B2201/054
• CPC Class Codes: B81B 3/0021 : Transducers for transforming electrical into mechanical energy or vice versa dynamo-electric machines H02K99/00; electrostatic machines H02N1/00; piezoelectric devices H10N30/00
• CPC Class Codes: B81C 1/00119 : Arrangement of basic structures like cavities or channels, e.g. suitable for microfluidic systems
• CPC Class Codes: C12Q 1/6846 : Common amplification features
• CPC Class Codes: G01N 27/447 : using electrophoresis
• 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 27/44791 : Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G05D 7/0694 : by action on throttling means or flow sources of very small size, e.g. microfluidics microvalves F16K99/0001; microstructural devices per se B81B
• CPC Class Codes: H01L 21/6715 : Apparatus for applying a liquid, a resin, an ink or the like H01L21/67126 takes precedence
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

Methods for amplifying nucleic acid, such as DNA or RNA, on a printed circuit board (PCB) substrate are disclosed. The amplification can be achieved through a variety of methods such as thermocycling or isothermally. The printed circuit board substrate can comprise an array of electrodes for transporting droplets and can be part of a sandwich structure including a top plate parallel to the printed circuit board substrate.

Claims

1. A method of amplifying DNA comprising:

(a) providing a sample solution comprising DNA on a printed circuit board substrate comprising an array of electrodes for transporting droplets on the substrate, and a dielectric coating atop the electrodes, wherein the printed circuit board substrate is provided as part of a sandwich structure, the sandwich structure further comprising;

(i) a top plate parallel to the printed circuit board substrate and forming a gap between the top plate and the printed circuit board substrate;

(ii) a seal between the top plate and the printed circuit board substrate; and

(iii) oil in the gap; and

(b) amplifying the DNA on the printed circuit board substrate, wherein the amplifying is achieved by thermocycling wherein the amplifying is accomplished in a droplet in the oil in the gap.