User manual

Droplet Digital
™
PCR Applications Guide | 89
Droplet Digital
™
PCR Tips, Assay Considerations, and Troubleshooting
Fig. 9.9. Good separation of positive and negative droplets in EvaGreen ddPCR reactions. A–B, combined
primer concentration between 75–250 nM is shown, with 100–200 nM being optimal; C, effect of EvaGreen ddPCR
reaction with 900 nM primers on increasing concentrations of DNA; D, decreasing the primer concentration to
100 nM produces a uniform cluster of negative droplets.
Channel 1 amplitudeChannel 1 amplitude Separation value, arbitrary units
0 50,000 100,000 150,000 200,000 250,000 300,000
Event number
0 40,000 80,000 120,000
Event number
0 20,000 40,000 60,000 80,000 100,000
Event number
0 50 100 150 200 250 300 350 400 450 500 550 600 650 700 750 800 850 900 950 1,000
Primer concentration, nM
1 µM 300 nM 25 nM
A07 B07 C08 D07 E07 F07 G07 H07 A09 B09 C09 D09 E09 F09 G09 H09 A11 B11 C11
A06 B06 C06 D06 E06 F06 G06 H06 B07 B08 D07 D09 F08 F09
100 nM increments 25 nM increments
40,000
35,000
30,000
25,000
20,000
15,000
10,000
5,000
0
25,000
20,000
15,000
10,000
5,000
0
25,000
20,000
15,000
10,000
5,000
0
22.5
20
17. 5
15
12.5
10
7.5
5
2.5
0
A
C D
B