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In experiments aimed at adjusting the number of precipitate washes needed, LDL precipitate was washed once or twice with precipitating solution, or not washed at all, prior to solubilizing.
Whole blood (0.1 mL) was added to distilled water (1.9 mL) together with 3 mL of precipitating solution (1.67 g glacial metaphosphoric acid, 0.2 g disodium ethylenediaminetetraacetic acid, EDTA, and 30 g sodium chloride).
The dialysate/carrier solution was further processed by adding 1.5 mL magnesium precipitating solution (6.05 g/L Trizma base, 5.85 g/L NaCl, and 100 g/L MgCl2·6H2O, pH 9.274); the solution was vortexed and allowed to stand for 10 min. The precipitate was centrifuged for 5 min at 2,000 rpm and the supernatant was removed.
Grape juice (commercially available from supermarket) was included as a control, and was diluted similarly, firstly with perchloric acid precipitating solution, and then serially diluted with phosphate buffer (75 mM, pH 7.4), giving final concentrations of 20, 10, 5, and 2.5 μL/mL.
Color detection was performed using 2 ml of BM purple precipitating solution (Roche).
After the Protein Precipitating Solution (PPS) was added to the lysate and the samples were centrifuged, 200 µl of sample was added to the QIAxtractor Instrument and purified using the QIAxtractor Liquid Sample Protocol.
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The precipitated solution was cooled down to 30 °C.
The MMV subjected to PCR (PCR MMV) was spun to precipitate solutions to the bottom before the following operation.
The metals were selectively recovered by precipitating the solution by adjusting the pH.
Crystals were cryoprotected in precipitating reservoir solution enriched in 10% glycerol before flash-freezing to 100 K.
Proteins were subsequently precipitated with Protein Precipitation Solution (Gentra Systems) and spun down.
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