technique
Aqueous based droplets are created in an oil environment, each droplet functions as a micro reaction chamber. In ddPCR the sample is partitioned over 20.000 droplets such that within each droplet a maximum of one target molecules are present, followed by PCR amplification of the template molecules, a fluorescent read-out of the individual droplet showing either no or a signal (‘digital output’) allows for the absolute quantification of the number of target molecules in the sample.
ddPCR technology uses a flexible chemistry and workflows similar to those used for most standard TaqMan probe based and EvaGreen assays. Applications can be found in somatic mutation and CNV detection, gene expression analysis, microorganism detection and accurate quantification and qualification of NGS libraries.
At the CFG researchers can have access to a QX200 ddPCR instrument of BioRad. The CFG provides the chemistry for probe detection, you only have to bring your own assay and your samples.
Photograph by Paulien Kluver