Systems and methods for performing digital pcr

Abstract

Systems and methods are described for quantifying a target nucleic acid. A sample comprising a target nucleic acid is segregated into a first plurality of the reaction volumes containing at least one target nucleic acid molecule and a second plurality of the reaction volumes containing no target nucleic acid molecules. The reaction volumes are subjected to an amplification assay, wherein the amplification assay is configured to amplify the target nucleic acid. An indicator of amplification is detected or measured in at least some of the plurality of reaction volumes. The target nucleic acid is quantified based on the detection or measurement. After discontinuing the amplification assay, the plurality of reaction volumes may be heated and changes in the indicators of amplification of two or more of the at least some of the reaction volumes may be detected or measured.

1 .- 39 . (canceled) 
     
     
         40 . A system for detecting or quantifying a nucleic acid in a sample, the system comprising:
 a reaction device;   an electronic processor;   a memory comprising instructions for performing steps comprising:
 segregating a sample comprising a target nucleic acid into a plurality of sample reaction volumes, wherein the plurality of sample reaction volumes include a first plurality of the sample reaction volumes each containing at least one molecule of the target nucleic acid and a second plurality of the sample reaction volumes each containing no molecules of the target nucleic acid; 
 subjecting the plurality of sample reaction volumes to an amplification assay, wherein the amplification assay is configured to amplify the target nucleic acid; 
 detecting or measuring indicators of amplification for at least some of the sample reaction volumes of the plurality of sample reaction volumes; 
 discontinuing the amplification assay; and 
 after discontinuing the amplification assay, heating the plurality of sample reaction volumes and detecting or measuring changes in the indicators of amplification for two or more of the at least some of the sample reaction volumes; and 
   an input/output device comprising an input device and a display;   wherein the electronic processor performs the instructions and displays an amount of the target nucleic acid on the display.   
     
     
         41 . The system of  claim 40 , wherein the memory is a random access memory. 
     
     
         42 . A system for detecting or quantifying a nucleic acid in a sample, the system comprising:
 an electronic processor;   a memory comprising instructions for performing steps comprising:
 segregating a sample comprising a target nucleic acid into a plurality of sample reaction volumes, wherein the plurality of sample reaction volumes include a first plurality of the sample reaction volumes each containing at least one molecule of the target nucleic acid and a second plurality of the sample reaction volumes each containing no molecules of the target nucleic acid; 
 subjecting the plurality of sample reaction volumes to an amplification assay, wherein the amplification assay is configured to amplify the target nucleic acid; 
 detecting or measuring indicators of amplification for at least some of the sample reaction volumes of the plurality of sample reaction volumes; 
 discontinuing the amplification assay; and 
 after discontinuing the amplification assay, heating the plurality of sample reaction volumes and detecting or measuring changes in the indicators of amplification for two or more of the at least some of the sample reaction volumes; and 
   an input/output device comprising an input device and a display;   wherein the electronic processor performs the instructions and displays information regarding a detection of the target nucleic acid on the display.   
     
     
         43 . The system of  claim 42 , wherein the memory is a random access memory. 
     
     
         44 . A system for detecting or quantifying a nucleic acid in a sample, the system comprising:
 a reaction device configured to contain or provide a plurality of sample reaction volumes from a sample solution containing a nucleic acid;   an electronic processor;   a temperature controller;   a memory comprising instructions for (1) detecting or measuring an indicator of amplification for at least some of the sample reaction volumes, (2) heating the plurality of sample reaction volumes, and (3) detecting or measuring a change in the indicator of amplification for one or more of the at least some of the sample reaction volumes;   an output device comprising a display;   wherein the electronic processor performs the instructions and displays at least one of (1) a detection and/or an amount of the indicator of amplification and/or (2) the change in the indicator of amplification of one or more of the at least some of the sample reaction volumes.   
     
     
         45 . The system of  claim 44 , wherein the memory is a random access memory. 
     
     
         46 . The method of claim  1 , wherein detecting or measuring indicators of amplification comprises detecting or measuring one indicator of amplification for each of the at least some of the sample reaction volumes.