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Step 3. Finally, smush some marshmallows into the gunk and blast your piggy for 20 mins in a hot oven.
The surface cells released by this treatment were collected by centrifugation (the supernatant generated by 1900 rpm for 5 mins in a bench top centrifuge).
Slides were heated for 40 mins in a staining dish containing sodium citrate buffer (10 mM citric acid, 0.05% Tween 20, pH 6.0) until the temperature reached 95 100°C, followed by cooling to room temperature.
PCR conditions: initial denaturation at 95°C for 1 min, followed by 15 cycles of denaturation at 95°C for 30 s, annealing at 45°C for 30 s and extension at 72°C for 30 s, and 15 cycles of 95°C for 15 s, 55°C for 30 s and 72°C for 30 s followed by a final extension at 72°C for 5 mins in a PCR thermal cycler (eppendorf).
The mixture was incubated at 37°C for 30 mins in a water bath.
Sections were incubated for 20 mins in a 0.6% BSA blocking solution and probed with the primary antibody (1 50 dilution) for 2 hours at room temperature.
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Detection from sample application to final results was completed in 6 min in a reagentless process.
The LAMP assay was performed at 62 °C for 60 min in a water bath.
Thereafter, the female's behavior was monitored for 20 min in a male preference procedure.
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