Method for chemical amplification based on fluid partitioning in an immiscible liquid

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.

The invention claimed is: 
     
       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 of nucleic acid amplification of a sample, comprising the steps of:
 partitioning the sample into partitioned sections,
 wherein the sample comprises a nucleic acid and components for performing nucleic acid amplification, 
 wherein the step of partitioning the sample into partitioned sections comprises flowing the sample through an injection orifice into an immiscible carrier fluid, 
 wherein the partitioned sections comprise a gelling agent or a gel, and 
   subjecting the partitioned sections of the sample to nucleic acid amplification.    
     
     
       12. The method of claim 11, wherein the injection orifice injects the sample and a PCR reagent.  
     
     
       13. The method of claim 11, wherein the injection orifice connects a sample flow pathway to a channel or tube comprising an immiscible fluid.  
     
     
       14. The method of claim 11, wherein the partitioned sections are separated by the immiscible carrier fluid.  
     
     
       15. The method of claim 11, wherein the nucleic acid amplification comprises amplification of a DNA target.  
     
     
       16. The method of claim 15, wherein the partitioned sections contain, on average, a single template of a DNA target, and wherein the single template is amplified within the partitioned sections.  
     
     
       17. The method of claim 15, wherein the partitioned sections are exposed to multiple heating and cooling cycles.  
     
     
       18. The method of claim 17, further comprising moving the partitioned sections through a microchannel.  
     
     
       19. The method of claim 11, further comprising detecting the products of the nucleic acid amplification.  
     
     
       20. The method of claim 19, wherein the detecting comprises confocal imaging.  
     
     
       21. The method of claim 19, wherein the detecting comprises laser excitation.  
     
     
       22. The method of claim 19, wherein the partitioned sections are probed for fluorescent signal.  
     
     
       23. The method of claim 11, wherein the partitioned sections comprise microdroplets or slugs.