twoddpcr

twoddpcr performs automated analysis of two-channel droplet digital PCR (ddPCR) data to classify droplets into four fluorescence-based clusters and quantify mutant and wild-type target molecules using Poisson statistics.

Key Features:

• Automatic Classification: Classifies droplets into four clusters based on two-channel fluorescence amplitudes to distinguish mutant-only, wild-type-only, both, and empty droplets.
• Poisson Statistics: Applies Poisson-distribution principles to estimate absolute target molecule counts from partitioned droplet data.
• QuantaSoft Data Import: Imports fluorescence amplitude measurements exported from Bio-Rad's QuantaSoft.
• Dual-Channel Allele Detection: Distinguishes and quantifies mutant and wild-type alleles within the same ddPCR experiment using two fluorescence channels.

Scientific Applications:

• Circulating Free Tumor DNA Analysis: Quantifies low-abundance mutant DNA in plasma for non-invasive cancer diagnostics and monitoring.
• Genetic Mutation Detection: Detects and quantifies mutant and wild-type alleles within complex samples.

Methodology:

Data are imported from Bio-Rad's QuantaSoft as fluorescence amplitude measurements, droplets are classified into four clusters based on two-channel fluorescence intensities corresponding to mutant-only, wild-type-only, both, or neither, and Poisson distribution principles are used to estimate absolute target molecule numbers per category.