User manual

Droplet Digital
™
PCR Applications Guide | 45
5 Rare Mutation and
Sequence Detection
Overview
Droplet Digital
™
PCR (ddPCR
™
) enables detection and analysis of nucleic acids at a level
of sensitivity and precision beyond the capabilities of previous methods. Applications that
focus on the lower limits of nucleic acid detection can be separated into two classes:
■
Rare mutation detection
■
Rare sequence detection
Rare mutation detection (RMD) and rare sequence detection (RSD) can be classified
according to their assay components. An RMD assay is made up of a single primer set
plus two competitive probes (Figure 5.1). RSD uses either a single assay to detect a rare
sequence per unit volume (Case 1) or two independent, noncompetitive assays to detect
a rare sequence as a proportion of a more common sequence (Case 2). Both RMD and
RSD take advantage of the high sensitivity of ddPCR.