User manual

Droplet Digital
™
PCR Applications Guide | 71
Next-Generation Sequencing Library Analysis
1,500 —
850 —
700 —
500 —
400 —
300 —
200 —
150 —
100 —
50 —
15 —
bp bp
Ladder AF NTC SM NTC AF 1.2 SM 1 AF NTC SM NTC AF 1.2 SM 1 AF NTC SM NTC AF 1.2 SM 1
L 1 2 3 4 5 6 7 8 9 10 11 12
— 1,500
— 850
— 700
— 500
— 400
— 300
— 200
— 150
— 100
— 50
— 15
Fig. 7.9. Postrecovery analysis of ddPCR products. bp, base pair; NTC, no template control.
In Figure 7.9 four replicate wells are shown for each assay, of which three were combined
for the droplet breaking protocol, and the fourth was read by the QX100 droplet reader.
Gel analysis and electropherograms of amplicon products made from two different
reactions prepared in droplets and no template controls are shown in the panels to the left.
Lanes 1, 2, 5, 6, 9, and 10 are no template controls of the reactions; lanes 3, 7, and 11
are a Staphylococcus aureus assay; and lanes 4, 8, and 12 are a different assay for the
S. aureus template.
Fig. 7.10. ddPCR quantification of droplet recovered amplicons. CPD, copies per droplet;
NTC, no template control; SA1, S. aureus assay 1; SA2, S. aureus assay 2.
SA1 NTC SA2 NTC SA1 1.2 CPD SA2 1.2 CPD
Sample
Concentration, copies/µl
2,000
1,600
1,20 0
800
400
0
1,670
1,500
0
0.0809
Results of ddPCR concentration plots for the two S. aureus reactions carried out in the
example are shown.