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It took about 2 min for final setting of the reaction pressure and reaction time will be considered as zero.
For concentrated nanofluids, stock solutions of gold nanoparticles synthetized by hydroquinone method were centrifuged at 6,000 rpm for 30 min for final concentrations of 1 mg/ml.
PCR parameters were 95 °C for 5 min of initial denaturation, 95 °C for 30 s of denaturation, 59 °C for 2 min of annealing, 72 °C 4 min and 72 °C of 10 min for final elongation on Eppendorf Mastercycler personal, Germany.
PCR was carried out in a PCR Sprint thermal cycler (Thermo electron corporation, Milford, MA) under the following conditions: initial denaturation at 95°C for 15 min followed by 35 cycles of 94°C for 1 min, 62°C for 1 min, 72°C for 1 min, followed by 72°C for 10 min for final extension.
RT-PCR reaction was carried out in a PCR Sprint thermal cycler (Thermo electron corporation, Milford, MA) under the following conditions: initial denaturation at 95°C for 15 min followed by 35 cycles at 94°C for 1 min, 60°C for 1 min, 72°C for 1 min, followed by 72°C for 10 min for final extension.
The mixture was maintained at 72°C, for 10 min, for final extension.
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The PCR conditions were: An initial step at 94 °C for 30 s, 15 cycles at 94 °C for 15 s, 62 °C for 30 s, and 70 °C for 15 s, followed by 75 °C for 5 mins for final extension.
In this case it was about 23 min for a final pH of 7.4.
The time per PET bed position was varied from 90 s for the first session to 4 min for the final session to compensate for radioactive decay.
Peptides were extracted twice more with 12 µl formic acid solution and 12 µl acetonitrile for 30 min for a final volume of 60 ml.
Thermal cycling was performed under the following conditions: 94°C for 5 min, 28 cycles at 94°C for 1 min, 54°C for 1 min, and 72°C for (1 min), followed by 72°C (5 min) for the final extension.
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