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The solvent was allowed to evaporate for 10 s at 20 cm and light cured for 10 s.
qPCR was performed with the Light Cycler 2.0 instrument (Roche) using the program: 95 °C for 2 min, 45 cycles of 95 °C for 10 s, 59 °C for 10 s, and 72 °C for 10 s.
The vestibular faces of all teeth were cleaned with a rubber cup and non-fluoridated pumice-water slurry (S.S. White, Petrópolis, RJ, Brazil) for 10 s, rinsed with air water spray for 10 s and air-dried for 10 s.
The electrode was held at −0.4 V for 10 s prior to cyclic voltammetry or −0.8 V for 10 s prior to linear sweep and square-wave voltammetry.
Fluorescence was monitored for 10 s after the last cycle.
Sonication was performed 6 times for 10 s each.
The PCR conditions were: 95°C for 10 min, followed by 40 cycles at 95°C for 10 s, 60°C for 10 s and 72°C for 10 s.
The conditions for PCR were denaturation at 95°C for 10 s, annealing at 62°C for 10 s, and elongation at 72°C for 14 s.
PCR conditions were standardized to 40 cycles; 95°C for 10 s, 55°C for 10 s, and 72°C for 30 s. Results were analyzed as described.
The samples were injected at a pressure of 5.0 kPa for 10 s (approximately 10 nL).
Animals were lysed in this buffer for 10 s using the Precellys 24 Lysis and Homogeniser.
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