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The pellet was resuspended in a small volume of ice-cold buffer containing 10 mM Tris-HCl (pH 7.6), 10% (v/v) glycerol, 1 mM DTT, and 1 mM EDTA.
Cells were then suspended in a small volume of ice-cold PBS, typically 200 – 400 µL depending on the estimated amount of cells isolated.
The final pellet was resuspended in a small volume of ice-cold distilled water and stored at 4°C until use.
Red cell lysis was performed by incubating the cellular pellet with a small volume of ice-cold distilled water for 20 s, followed by quenching with a large volume of isotonic PBS.
The ELP pellets were dissolved in a tenfold smaller volume of ice-cold Milli-Q water and centrifuged at 4°C for 45 min and 15,000 rpm in an SS34 rotor.
The results indicate that we should pay more attention to possibly small volume of PCR products in ice bath and minor temperature difference of two liquids in operation.
A small volume of intermediate lava was erupted from Ice Peak compared to the other central volcanoes.
1C11 adherent cells were washed twice in PBS then scraped on ice in a small volume of PBS containing a cocktail of protease inhibitors (Complete™ from Roche) and 1 mM sodium orthovanadate (Na3VO4) phosphatase inhibitor.
On the next day, vesicles were re-suspended in a small volume of the same buffer and then kept on ice until immediately prior to transport measurements when samples were warmed to 37°C.
The pelleted bacteria were placed on ice and resuspended in a small volume of 270 mM sucrose.
The frozen cells were suspended in a small volume (0.2 ml) of ice-cold 0.1 M Tris-HCl buffer, pH 7.5, containing 0.15 M NaCl, 1% Triton X-100, 0.5% sodium deoxycholate, and a protease inhibitor mixture (2 μg antipain/ml, 0.1% aprotinin, 10 μg benzamidine/ml, 1 μg chymostatin/ml, 1 μg leupeptin/ml, and 1 μg pepstatin/ml) as described (17).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com