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. Author manuscript; available in PMC: 2015 Jul 1.
Published in final edited form as: Hum Pathol. 2014 Apr 13;45(7):1546–1550. doi: 10.1016/j.humpath.2014.03.013

Figure 2.

Figure 2

Schematic of ddPCR workflow for assessing LOH. A) The ddPCR reaction involves combining fragmented DNA to be studied with droplet oil and PCR reagents (PCR master mix, primers and probes). The droplet generator machine partitions the required reagents and DNA fragments into approximately 2 × 104 individual droplets per well. Following droplet generation, end-point PCR is carried out on a standard thermo-cycler. B) After PCR amplification and generation of unquenched fluorophores, the droplets are read on a droplet reader. Example #1 illustrates the expected distribution of fluorescent droplets for no LOH, where 50% of the gated droplets are positive for the wild-type allele and 50% are positive for the VUS. Example #2 illustrates the approximate distribution of fluorescent droplets for LOH (with loss of the wild-type allele) with contaminating wild type from surrounding normal tissue.