Method for chemical amplification based on fluid partitioning in an immiscible liquid
A system for nucleic acid amplification of a sample comprises partitioning the sample into partitioned sections and performing PCR on the partitioned sections of the sample. Another embodiment of the invention provides a system for nucleic acid amplification and detection of a sample comprising partitioning the sample into partitioned sections, performing PCR on the partitioned sections of the sample, and detecting and analyzing the partitioned sections of the sample.
1. An apparatus for nucleic acid amplification of a sample, comprising:
means for partitioning said sample into partitioned sections, wherein said means for partitioning said sample into partitioned sections comprises an injection orifice, and
means for performing PCR on said partitioned sections of said sample.
2. The apparatus for nucleic acid amplification of a sample of claim 1 wherein said injection orifice is an injection orifice that produces microdroplets.
3. The apparatus for nucleic acid amplification of a sample of claim 1 wherein said injection orifice is an injection orifice that injects said sample and a PCR reagent.
4. The apparatus for nucleic acid amplification of a sample of claim 1 wherein said means for performing PCR on said partitioned sections of said sample comprises a continuous tube for circulating said partitioned sections of said sample through a heater to perform PCR.
5. The apparatus for nucleic acid amplification of a sample of claim 1 wherein said means for performing PCR on said partitioned sections of said sample comprises a continuous tube for circulating said partitioned sections of said sample through a heater and cooler to perform PCR.
6. The apparatus for nucleic acid amplification of a sample of claim 1 wherein said means for performing PCR on said partitioned sections of said sample comprises a pump, a continuous tube, and a heater.
7. The apparatus for nucleic acid amplification of a sample of claim 1 including means for detection and analysis of said partitioned sections of said sample comprising a laser and a detector.
8. The apparatus for nucleic acid amplification of a sample of claim 1 including means for detection and analysis of said partitioned sections of said sample comprising a blue laser and a detector.
9. The apparatus for nucleic acid amplification of a sample of claim 1 wherein said means for partitioning said sample into partitioned sections comprises means for separating said sample into immiscible slugs.
10. A method of nucleic acid amplification of a sample, comprising the steps of:
partitioning said sample into partitioned sections, wherein said step of partitioning said sample into partitioned sections comprises flowing said sample through an injection orifice, and
subjecting said partitioned sections of said sample to PCR.
11. A method for amplifying nucleic acid, the method comprising:
contacting an aqueous solution comprising nucleic acids and components for performing nucleic acid amplification with an immiscible fluid thereby partitioning the aqueous solution into microdroplets comprising nucleic acids and components for performing nucleic acid amplification;
arranging the microdroplets in small indentations in a two dimensional array on a solid substrate support; and
thermocycling the microdroplets in the two dimensional array to amplify the nucleic acids.
12. The method according to claim 11, wherein the small indentations are microwells.
13. The method according to claim 11, further comprising detecting products of the nucleic acid amplification.
14. The method according to claim 13, wherein detecting is optically detecting.
15. The method according to claim 14, wherein optically detecting comprises scanning each microdroplet.
16. The method according to claim 14, wherein optically detecting comprises use of a confocal imaging system.
17. The method according to claim 14, wherein optically detecting is fluorescently detecting.
18. The method according to claim 17, wherein an amount of amplification is indicated by fluorescence.
19. The method of claim 11, wherein the solid substrate is hydrophobic.
20. The method of claim 11, wherein each microdroplet has a volume in the picoliter range.
21. The method of claim 11, comprising using a pump for contacting the aqueous solution with the immiscible fluid.
22. The method of claim 11, comprising using a pump for arranging the microdroplets in small indentations in the two dimensional array on the solid substrate support.
23. A method for amplifying nucleic acid, the method comprising:
contacting an aqueous solution comprising nucleic acids and components for performing nucleic acid amplification with an immiscible fluid thereby partitioning the aqueous solution into microdroplets comprising nucleic acids and components for performing nucleic acid amplification, wherein each microdroplet has a volume in the picoliter range;
arranging the microdroplets in small indentations in a two dimensional array on a hydrophobic solid substrate support;
thermocycling the microdroplets in the two dimensional array to amplify the nucleic acids; and
optically detecting nucleic acid amplification products via fluorescence or absorption.