Partition-based method of analysis
Methods of partition-based analysis. In an exemplary method, a device having a port fluidically connected to a chamber may be selected. A sample-containing fluid may be placed into the port. The sample-containing fluid may be moved from the port to the chamber. Partitions of the sample-containing fluid may be formed. A monolayer of the partitions in the chamber may be created. At least a portion of the monolayer may be imaged.
1. A method for quality control in digital PCR, the method comprising the steps of:
preparing a reaction mixture comprising a target nucleic acid, a first dye, a second dye, and nucleic acid amplification reagents;
generating a plurality of partitions comprising the reaction mixture; and
assessing the first dye or and the second dye to indicate variation among the partitions, the first dye providing a test signal and the second dye providing a reference signal and indicating a partition volume of the partitions.
2. The method of claim 1 , wherein the assessing step comprises colorimetric measurement of the first dye or the second dye.
3. The method of claim 1 , wherein at least one of the partitions does not contain the reaction mixture.
4. The method of claim 1 , wherein the assessing step comprises assessing a partition volume for members of the plurality.
5. The method of claim 1 , further comprising the step of identifying a member of the plurality below a predefined partition volume.
6. The method of claim 1 , wherein reference signal indicates amplification of a control nucleic acid target.
7. A method for quality control in a digital PCR, the method comprising:
distributing a reaction mixture comprising a target nucleic acid, a first dye, a second dye, and nucleic acid amplification reagents into a plurality of partitions; and
assessing the first dye or and the second dye to indicate presences of the reaction mixture within members of the plurality, the first dye providing a test signal and the second dye providing a reference signal and indicating a partition volume of the partitions.
8. The method of claim 7 , wherein the assessing step comprises colorimetric measurement of the first dye or the second dye.
9. The method of claim 7 , wherein at least one of the partitions does not contain the reaction mixture.
10. The method of claim 7 , wherein the dye or the second dye is indicative of a volume of the reaction mixture in members of the plurality.
11. The method of claim 7 , wherein the assessing step comprises assessing a partition volume for members of the plurality.
12. The method of claim 7 , further comprising the step of identifying a member of the plurality of partitions below a predetermined partition volume.
13. The method of claim 7 , wherein the reference signal indicates amplification of a control nucleic acid target.
14. The method of claim 1 , wherein the dye or the second dye is indicative of a volume of the reaction mixture in members of the plurality.
15. The method of claim 1 , wherein the second dye is not coupled to an amplification reaction and provides a passive reference.
16. The method of claim 1 , further comprising transforming the test signal based on the reference signal to reduce variation.
17. The method of claim 7 , wherein the second dye is not coupled to an amplification reaction and provides a passive reference.
18. The method of claim 7 , further comprising transforming the test signal based on the reference signal to reduce variation.