Methods and systems for volume variation modeling in digital PCR

Abstract

A method for performing digital polymerase chain reaction (dPCR) is provided. The method includes partitioning a biological sample volume including a plurality of target nucleic acids into a plurality of partitions, where at least one partition includes at least one target nucleic acid. The method further includes determining a model for volume variation of the plurality of partitions and determining a number of partitions including at least one target nucleic acid. The method includes generating a concentration of target nucleic acids in the biological sample based on the model for volume variation and the fraction of partitions including at least one target nucleic acid.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims the benefit of priority of U.S. Provisional Patent Application No. 62 / 152,542, filed on Apr. 24, 2015, and U.S. Provisional Patent Application No. 62 / 193,932, filed on Jul. 17, 2015, which are both incorporated herein in their entireties by reference.BACKGROUND[0002]The great promise of digital PCR is the potential for unparalleled precision enabling accurate measurements for genetic quantification. Target nucleic acid molecules of a sample requiring quantification are distributed evenly on a digital PCR consumable with many partitions and subjected to a PCR reaction. Partitions with template show amplification of the target nucleic acid and partitions lacking template do not show amplification. The observations are typically fitted with the Poisson model to predict the number of molecules present in the sample under measurement.[0003]A basic assumption of the Poisson model is that molecules are equally likely to b...

AI Technical Summary

Problems solved by technology

As positives may contain more than one copy of the target molecule, a simple summing of the number of positives will not yield the correct number of target molecules present across the partitions.

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