Compositions and methods for digital polymerase chain reaction

Abstract

At some aspects, the present disclosure provides methods for identifying sequence variants in a nucleic acid sample. In some embodiments, a method comprises distinguishing between a true mutation in a polynucleotide and a random error introduced during an amplification step. In some embodiments, the methods reduce the number of false positives reported by a digital PCR assay. In some embodiments, the methods improve the accuracy of a digital PCR assay.

Description

[0001] cross reference [0002] This application claims the benefit of U.S. Provisional Application No. 62 / 694,324, filed July 5, 2018, which is hereby incorporated by reference in its entirety. Background technique [0003] Digital polymerase chain reaction (PCR) is a modification of traditional PCR methods that allows users to directly quantify nucleic acids in samples. Digital PCR methods typically involve partitioning the sample into discrete partitions such that each partition can be interrogated individually. Digital PCR is very sensitive but can be difficult to scale due to the limited number of pathways (plexes) that can be assayed in one reaction. This issue can be even more problematic for liquid biopsies that use cell-free DNA (cfDNA) as input, as starting material is often scarce. One way to address this issue might be to amplify cfDNA prior to performing digital PCR in order to provide sufficient starting material to separate into different assays. However, er...

AI Technical Summary

Problems solved by technology

However, errors introduced during the amplification step can generate false positives by digital PCR

This can be challenging for low allele frequency variant detection

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