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The plate was incubated on a shaking platform at room temperature for 30 minutes, and then overnight at 4°C.
Media was aspirated and cells were rinsed with PBS and fixed with 0.1% glutaraldehyde in PBS for 15 minutes on a rocking platform at room temperature.
Individual arrays were blocked with 1% blocking-grade blocker (Bio-Rad) in PBS for 1.25 hours on a rocking platform at room temperature.
This was followed by coomassie staining for 6 h in shaking platform at room temperature, and de-staining (40%% methanol and 10%% glacial acetic acid) for 2 h at room temperature.
Right colons were washed with shaking in 1× PBS buffer + 1 mM dithiothreitol (DTT) and then transferred into EDTA buffer (1 mM EDTA in PBS + 0.5 mM DTT) and placed on a shaking platform at room temperature.
The second part of the endometrial biopsy was immediately placed in RNAlater (Life Technologies Australia, Mulgrave, Vic, Australia), incubated overnight on a rocking platform at room temperature, then stored at −80 °C until further processed.
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Immunostaining was performed on the Dako Autostainer platform (DakoCytomation) at room temperature.
The plate was incubated for 2 h on a platform rotator at room temperature.
Asci were treated with 0.5% glusulase (Perkin Elmer) solution and incubated on a rotating platform overnight at room temperature.
Powdered material (2 g) was extracted with acetone (20 ml, Merck technical grade) using a platform shaker (Labotec) at room temperature for 30 min [ 26].
To isolate the soluble and insoluble fractions, cell pellets (cell fraction, C) from 0.8 mL of culture were resuspended by 0.5 mL of BugBuster Protein Extraction Reagent and incubated on a shaking platform for 10 20 min at room temperature.
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