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Sample clean-up was achieved by adding acetonitrile for protein precipitation.
Gel pieces were dehydrated and washed at 40°C by adding acetonitrile for 5 min. Gel pieces were dried for 5 min at 40°C and then re-hydrated at 4°C for 40 min with the trypsin solution (6 ng/µl of sequence grade trypsin in 25 mM ammonium bicarbonate).
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After adding acetonitrile (350 μl), the mixture was shaken for 30 seconds ×2, and centrifuged at 20,000 g, 4°C, for 10 minutes for two cycles.
Dehydrated gel pieces were produced by adding acetonitrile (100 μl), and then submitting them to air-drying for 5 10 min.
Upon adding acetonitrile into ionic liquid, the EDL capacitance curve switches from bell-shaped for neat IL to camel-shaped with 50 mol% and 95 mol% acetonitrile, and the EDL capacitance increases significantly at higher cathodic and anodic polarizations.
For plasma and homogenate samples, the proteins were precipitated by adding acetonitrile at a ratio of 1 3.
The gel pieces were crushed, dehydrated by adding acetonitrile, rehydrated by adding 10 20 µl of 25 mM ammonium bicarbonate with 10 ng/µl of sequencing grade trypsin (Promega), and incubated at 37°C for 15 17 h.
The method was repeated by adding 100% acetonitrile for 15 minutes.
This paper reports the work to find the best additives added to acetonitrile (ACN) for improving the separation of C4 mixtures with the help of computer-aided molecular design (CAMD).
The extracts were placed back into the Nalgene jars and 20 mL acetonitrile was added, homogenized for 2 min and the dispersing element rinsed with 5 mL acetonitrile.
We precipitated the proteins by adding 150 μL acetonitrile and vigorously shaking for 30 min, and afterward we centrifuged the samples and transferred them to autosampler vials.
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