Supplementary material
Figure S1 A) The temperature is controlled via a dual thermal controller setup. The thermoelectric
pump on the left can heat the sample (red) while the pump on the right maintains the rest of the chip at
a lower temperature (blue). B) The first thermoelectric element is used to heat the incubation region
(highlighted in red) while the second maintains the rest of the chip at room temperature (blue) to
prevent evaporation of the wash and desulfonation buffers during all heating steps. The thermoelectric
elements are controlled by a proportional-integral-derivative (PID) controller, which monitors the
heating surface temperature via a thermistor. The use of two temperature zones ensures that reagent
temperatures at incubation zones do not adversely affect other reagents due to thermal conduction.
Figure S2 Evaluation of droplet size after incubation. The droplet showed a 1% reduction after the 8
minutes of heating at 95 ˚C and a 0.5 % reduction after an hour at 60 ˚C, for a total of 1.5% volume
reduction.
Bisulfite conversion reagents:
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
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M-Binding buffer (Zymo D5001-3)
Lightning Conversion Reagent (Zymo D5030-1)
L-Desµlphonation buffer (Zymo D5030-5)
M-Wash buffer (Zymo D5001-4)
M-Elution buffer (Zymo D5001-6)
MagneSil® Red (Promega A1641)
Droplet volumes
I.
II.
III.
IV.
V.
VI.
VII.
Sµlfonation + Deamination
13 µl Lightning reagent + 2 µl DNA + 5 µl Paramagnetic particles
Binding
60 µl M-Binding Buffer
Wash
40 µl M-Wash Buffer
Desµlfonation
20 µl L-Desµlphonation Buffer
Wash
40 µl M-Wash buffer
Wash
40 µl M-Wash buffer
Elution
10 µl Elution Buffer