A 3-part CPD accredited course presented by Dr Jonathan Weiss Assistant Director of R&D at Qiagen. 

The first part of the course covers a general introduction to digital PCR and how it utilises partitioning and poison distribution to calculate target gene absolute copy numbers. Then the dPCR technologies including plate-based platforms and emulsion-based or ddPCR technologies. Like conventional QPCR a fluorophore and quencher molecular is typically against a target of interest although it also supports the use of intercalating DNA binding dyes. Following a run quality is assessed using 1D plots of the probe positive vs probe negative partitions, there should be a wide divergence. For multiple flurophores, a 2D scatter plot is used similar to flow cytometry. The technique is typically utilised for mutation detection and gene expression as with conventional QPCR and also for determining copy number variation.

Part two covers assay optimisation, with suboptimal assays a distinct divergence in the positive partitions is observed known as rain making it difficult to set a correct threshold. This can be overcome by using assays already by QPCR or by following quality guidelines such as ensuring they are less than 150bp, checking them on BLAST for off-target amplifications and also by adjusting the time, temperature and no. of cycles for amplification. 

Part three covers the usage of DPCR in environmental monitoring of wastewater, food, high throughput screening of GMOs and vector-borne disease detection as it detects even a single copy number it is highly sensitive and because of the use of partitions, it can be used to screen large pools of input material such as wastewater and crops for mutations selection.