Apparatus and methods for parallel processing of micro-volume liquid reactions

• US 7,833,719 B2
• Filed: 12/20/2007
• Issued: 11/16/2010
• Est. Priority Date: 02/18/2000
• Status: Expired due to Fees

• Current Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford Management Co.)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford Management Co.)
• Inventors: Foreman, Pamela K., O'Keefe, Matthew
• Primary Examiner(s): Beisner; William H
• Application Number:
  US12/004,116
• Publication Number:
  US 20080108112A1
• Time in Patent Office: 1,062 Days
• Field of Search: 435/288.4, 435/288.7, 435/303.1
• US Class Current: 435/6.1
• CPC Class Codes: B01F 2215/0431 Numerical size values, e.g. diameter of a hole or conduit, area, volume, length, width, or ratios thereof
• CPC Class Codes: B01F 33/30 Micromixers
• 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 33/30351 using hydrophilic/hydrophobic surfaces
• CPC Class Codes: B01F 35/71 Feed mechanisms with proportioning B01F35/80
• CPC Class Codes: B01F 35/712 for feeding fluids
• CPC Class Codes: B01F 35/7172 using capillary forces
• CPC Class Codes: B01J 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/0072 Organic compounds
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/0074 Biological products
• CPC Class Codes: B01L 2200/0642 Filling fluids into wells by specific techniques
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/10 Means to control humidity and/or other gases
• CPC Class Codes: B01L 2300/165 Specific details about hydrophobic, oleophobic surfaces
• CPC Class Codes: B01L 2400/0688 surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/50857 : using arrays or bundles of open capillaries for holding samples
• CPC Class Codes: B01L 3/5088 : confining liquids at a location by surface tension, e.g. virtual wells on plates, wires B01L3/50857 takes precedence
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: H01J 49/0418 : for laser desorption, e.g. matrix-assisted laser desorption/ionisation [MALDI] plates or surface enhanced laser desorption/ionisation [SELDI] plates
• CPC Class Codes: Y02P 20/582 : Recycling of unreacted starting or intermediate materials

Abstract

Disclosed herein are apparatuses and methods for conducting multiple simultaneous micro-volume chemical and biochemical reactions in an array format. In one embodiment, the format comprises an array of microholes in a substrate. Besides serving as an ordered array of sample chambers allowing the performance of multiple parallel reactions, the arrays can be used for reagent storage and transfer, library display, reagent synthesis, assembly of multiple identical reactions, dilution and desalting. Use of the arrays facilitates optical analysis of reactions, and allows optical analysis to be conducted in real time. Included within the invention are kits comprising a microhole apparatus and a reaction component of the method(s) to be carried out in the apparatus.

Claims

1. A method for simultaneously conducting a plurality of micro-volume polynucleotide amplification reactions, the method comprising:

(a) introducing a plurality of liquid samples into the sample chambers of a microhole apparatus, whereby the chambers contain necessary polynucleotide amplification reaction components;

(b) protecting the samples from evaporation by submerging the microhole apparatus in a hydrophobic medium during the plurality of micro-volume polynucleotide amplification reactions; and

(c) placing the apparatus into an environment favorable to the polynucleotide amplification reaction;

wherein the microhole apparatus comprises a substrate, wherein the substrate is solid and defines a plurality of sample chambers, wherein each sample chamber;

(i) extends through the substrate;

(ii) comprises one or more walls and an opening at each end; and

(iii) holds a sample such that the sample is retained in the apparatus through surface tension and such that (i) a liquid sample present in one sample chamber does not intermix with a liquid sample present in another sample chamber and (ii) the liquid samples are maintained in the sample chambers when the microhole device is submerged in a hydrophobic medium for a polynucleotide amplification reaction.

2. A method according to claim 1, wherein the polynucleotide amplification is a polymerase chain reaction.

3. A method according to claim 1, wherein the microhole apparatus is a titanium substrate.

4. A method according to claim 1, wherein the hydrophobic medium is an oil.

5. A method according to claim 4, wherein the oil is mineral oil.

6. A method according to claim 4, wherein the oil is silicone oil.

7. A method according to claim 1, wherein the micro-volume polynucleotide amplification reactions are optically monitored.