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To avoid evaporation of the medium, the plates were covered with a gas-permeable sealing membrane (Breathe-Easy, Diversified Biotech, Boston, USA).
After removal of uptake medium, the plates were immediately placed on ice and cells were lysed in ice-cold lysis buffer.
After absorption of drops in the agar medium the plates were sealed in plastic bags and incubated overnight at 37°C.
After adding the virus-containing medium the plates were centrifuged at 1500 rpm for 45 min at RT and subsequently cultured at 32°C and in 5% CO2 in the incubator.
After replacing the culture medium with the oxygen-glucose deprived medium the plates were transferred into an airtight pressure chamber (volume = 750 ml) equipped with inlet and outlet valves.
Similar(55)
The LVs was added to complete DMEM medium containing 8 μg/mL polybrene (Sigma-Aldrich) and exchanged with the medium in the plates.
Ten microliters of MTT solution (final concentration of 0.5 mg/mL) was added to the culture medium, and the plates were incubated for 4 h at 37 °C.
A. tumefaciens was co-cultivated with V. albo-atrum on and within tobacco stems in the absence of any medium on the plates.
Cells were plated after sonication on solid YPD medium and the plates were incubated at 25°.
Then, after removing the supernatant (lysate), the concentrate is plated on agar medium and the plates incubated overnight before counting.
For refreshing, new culture plates were filled with medium, and the plates were pre-warmed and oxygenated.
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