# US RE45,539 E1 · Method for chemical amplification based on fluid partitioning in an immiscible liquid > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US RE45,539 E1 - Document ID: USRE45539E1 - Status: Granted - Kind: E1 - Application number: 13/436,693 - Filed: 2012-03-30 - Granted: 2015-06-02 - Earliest claimed: 2003-03-14 - Assignees: Lawrence Livermore National Security, LLC - Applicants: Brian L. Anderson; Bill W. Colston; Christopher J. Elkin - Inventors: Brian L. Anderson; Bill W. Colston; Christopher J. Elkin - Web page: https://patentgrep.com/patent/USRE45539E1 ## CPC - C12Q 1/6844: Nucleic acid amplification reactions - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES ## Abstract 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. ## Claims ### Claim 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. ### Claim 2 (depends on 1) The apparatus for nucleic acid amplification of a sample of claim 1 wherein said injection orifice is an injection orifice that produces microdroplets. ### Claim 3 (depends on 1) 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. ### Claim 4 (depends on 1) 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. ### Claim 5 (depends on 1) 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. ### Claim 6 (depends on 1) 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. ### Claim 7 (depends on 1) 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. ### Claim 8 (depends on 1) 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. ### Claim 9 (depends on 1) 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. ### Claim 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. ### Claim 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. ### Claim 12 (depends on 11) The method according to claim 11, wherein the small indentations are microwells. ### Claim 13 (depends on 11) The method according to claim 11, further comprising detecting products of the nucleic acid amplification. ### Claim 14 (depends on 13) The method according to claim 13, wherein detecting is optically detecting. ### Claim 15 (depends on 14) The method according to claim 14, wherein optically detecting comprises scanning each microdroplet. ### Claim 16 (depends on 14) The method according to claim 14, wherein optically detecting comprises use of a confocal imaging system. ### Claim 17 (depends on 14) The method according to claim 14, wherein optically detecting is fluorescently detecting. ### Claim 18 (depends on 17) The method according to claim 17, wherein an amount of amplification is indicated by fluorescence. ### Claim 19 (depends on 11) The method of claim 11, wherein the solid substrate is hydrophobic. ### Claim 20 (depends on 11) The method of claim 11, wherein each microdroplet has a volume in the picoliter range. ### Claim 21 (depends on 11) The method of claim 11, comprising using a pump for contacting the aqueous solution with the immiscible fluid. ### Claim 22 (depends on 11) 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. ### Claim 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.