User manual

26 | Droplet Digital
™
PCR Applications Guide
Designing Droplet Digital
™
PCR Experiments
Fig. 2.14. Splice variant discrimination using the QX200 ddPCR EvaGreen supermix with a
single set of primer pairs.
EvaGreen and Gene Expression
Measuring gene expression using ddPCR and EvaGreen allows you to detect and quantify
splice variants as well as contaminating genomic DNA. Figure 2.14 shows an experiment
demonstrating that splice variants can be seen as a cluster of droplets with higher
fluorescence amplitude, referred to as a super cluster. Control samples with no reverse
transcriptase added were run to verify that the super cluster was not due to residual
genomic DNA.
35,000
30,000
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20,000
15,000
10,000
5,000
0
Channel 1 amplitude
F07 F08 F09 G07 G08 G09 H07 H08 H09
0 20,000 40,000 60,000 80,000 100,000 120,000
No template No reverse cDNA
control transcription
Event number
Super clusters