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The residue was subsequently extracted by shaking for 1 h with a 1 10 w/v of 0.1 mol/L NaOH and Na4P2O7 (1:1 v/v, pH = 13.0) solution and then the mixture was remained for 24 h.
Yolk was diluted ten times (v/v) with acidified water (pH 5.2), the mixture was remained 16 h at 4°C and, then, centrifuged at 8,500xg for 30 min at 4°C.
The samples were extracted by shaking for 1 h with a 1 10 w/v of 0.5 M Na2SO4 solution and then the mixture was remained for 24 h.
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But even if I'd beaten it with an airplane propeller, the mixture was determined to remain loose and glistening, so I gave up and poured the caramelized sugar onto a greased baking sheet.
Finally, the remaining mixture was added.
The remaining mixture was centrifuged at 35,860 × g for 1 h, and then, the suspended solution was removed.
The remaining mixture was placed in an alumina crucible and fired at 500 for 1 h in air.
A 3 mL dichloromethane/hexane/(1 1, v/v) mixture was added to the remaining solution after protein precipitation and the solution was vortex mixed for 1 minute and then centrifuged at 3500 g for 5 minutes at 4°C.
The remaining mixture was added with water to a final volume of 100 ml.
The same amount of the solvent mixture was added to the remaining pelleted biomass and the extraction procedure was repeated.
The plasmid DNA/CaCl2/HBS mixture was combined with the remaining medium and transferred back into the HYPERFlask vessel.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com