User manual

Additional Applications
76 | Droplet Digital
™
PCR Applications Guide
Fig. 8.5. Dilution series of an assay targeting chromosome 10 of the human genome.
Channel 2 amplitude
0
0
20,000
4,000
3,000
2,000
1,000
Event number
6,000
5,000
7,000
40,000 60,000 80,000 100,000
A02
Stock
B02
1/2
C02
1/4
D02
1/8
E02
1/16
F02
1/32
G02
1/64
H02
No assay
A good starting point would be to halve, or conversely, double the concentration of an assay
relative to the standard final 1x concentration in order to spatially resolve the two assays in a
2-D plot of FAM and HEX amplitudes.
Figure 8.5 is an assay targeting chromosome 10 of the human genome. The stock has the
highest fluorescence amplitude and subsequent twofold changes in starting concentration
demonstrate resolvable droplet populations based on fluorescence amplitude
down to one-quarter dilution.
Figure 8.6 represents an example of such a multiplexing strategy being used with
one FAM channel assay and two HEX channel assays in a 2-D plot of the data.
Fig. 8.6. 2-D plot of FAM amplitudes (channel 1) vs. HEX amplitudes (channel 2) for
a multiplex experiment.
Channel 1 amplitude
0
0
2,000
15,000
5,000
Channel 2 amplitude
4,000 6,000 8,000 10,000
Chr10q1 +
Chr13q3
Chr13q3
20,000
25,000
30,000
10,000
Chr10q1
RPP30
12,000