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US 2020/0370097 A1Published application
Quantitating high titer samples by digital pcr
OwnerJohn M. Clemens, Eric B. Shaininventors
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Abstract
The present invention provides systems, devices, methods, kits, and compositions for nucleic acid analysis using digital PCR. In particular, methods are provided to analyze high titer samples that cannot be divided into a sufficient number of partitions containing zero nucleic acid molecules per partition to allow for Poisson analysis (digital PCR analysis).
Claims20
A system, comprising:
- a) a reaction mixture, comprising:
- i) at least one target nucleic acid;
- ii) at least one internal standard nucleic acid at a concentration sufficient to produce a number of partitions containing zero copies of said internal standard nucleic acid per partition wherein λIS≈0.01-1, wherein λIS is the average number of internal standard copies per partition to allow for Poisson analysis, wherein said internal standard nucleic acid has at least one identical primer binding sequence as said target nucleic acid;
- iii) at least one primer oligonucleotide specific for said target nucleic acid and for said internal standard nucleic acid;
- iv) at least one amplification reagent;
- b) a partitioning device;
- c) an amplification device;
- and
- d) a detection device.
6 more elements
Dependencies are inferred from the claim text.