User's Manual

42
Chapter 6: Evaluation of Amplification
and Target Enrichment
Quantify total DNA
After dMDA incubation, break the dMDA droplets with Break solution ● and Break colour ● (Fig. 6.1).
1. Add 20 μl Break solution ● to each tube.
2. Add 1 μl of Break colour ●. This will colour the water phase. If colouring is too weak, add 1 extra μl.
Note: The water phase may be a colour ranging from yellow to purple as the Break colour is
functioning as a pH indicator as well.
3. Flick tube gently, do not vortex.
4. Spin tube briefly (15-30 sec).
5. Remove the clear Break solution phase from the bottom of the tube and discard.
6. Repeat steps 3-5 to remove all leftover Break solution. It is important to remove all the Break
solution as residual Break solution may inhibit downstream enzymatic reactions.
7. Keep the coloured water phase, which will contain the amplified DNA (Fig. 6.1).
Fig. 6.1. Break post incubation dMDA droplets with Break solution ● and Break colour ●. Discard the clear
break solution phase at the bottom of the tube. Keep the top coloured water phase, this phase will contain
your DNA molecules.
Measure the total amount of enriched DNA by a reliable and sensitive method such as with Qubit™,
Bioanalyzer™, TapeStation™, FEMTO Pulse™ or similar. If possible, evaluate the size of sorted and amplified
DNA fragments.
Coloured water phase (keep)
Break solution phase (discard)