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Amniotic fluid samples (8 12 ml) were spun at 4500 rpm for 30 minutes.
Thawed serum samples (10 mL) were spun at 16000 g for 60 min.
Treated cells (30 ml) were spun and resuspended in 20% trichloroacetic acid (TCA) for protein extract preparation at 0, 5, 10, 15 and 20 min after treatment.
The culture was cooled on ice for 30 minutes and then two lots of 500 ml were spun down at 4000 × g for 10 minutes at 4°C.
DT40 cell suspension cultures (∼5 × 10 cells per 0.1 ml) were spun onto slides at 1000 r.p.m. for 3 min using a cell spin cytocentrigfuge from Tharmac.
The culture was cooled down on ice for 30 min. Two lots of 50 ml were spun down at 3,000 rpm (Sorval) for 15 mins at 4°C, rinsed twice with 10 ml ice cold water, washed (with resuspension) with ice cold 10% glycerol and finally resuspended in 250 μl (per 50 ml of bacterial culture) of ice cold 10% glycerol.
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Several aliquots of each extract (0.1 mL, 3×0.2 mL and 0.4 mL) were spin-dried under vacuum and stored at −80°C until analysis.
The ionic PF in acetonitrile (1 mL/1 mL) was spin-coated on top of the TiO2 layer, followed by drying in a vacuum oven at 90 °C for 30 min.
For genomic DNA extraction and analysis of copy number, cell samples (∼40 ml each) were spun down, quickly chilled in liquid nitrogen and stored at −80°C for further analysis.
Samples were taken at 5-min intervals for a period of 60 min. For DNA content analysis, cell samples (∼5 ml each) were spun down, resuspended in ice-cold 70% ethanol and stored at 4°C for subsequent analysis.
In addition, SSE-treated cells (1 × 10 cells/0.2 mL PBS) were spun onto glass slides by cytospin centrifuge (Cellspin; Hanil, Korea) at 400 × g for 4 min, fixed with 4% paraformaldehyde for 10 min at 37°C, stained with DAPI solution for 10 min, and then observed under the fluorescence microscope (Olympus TH4-200; Optical OptiCo. Co. LTD).
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