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In the last step of QCM measurements, the immobilization buffer was passed through the apparatus chamber (step 7, Fig. 1).
At first (step 1, Fig. 1), the immobilization buffer was passed through the chamber until the baseline frequency was constant.
To obtain a baseline for investigation of DNA uranyl interactions, the immobilization buffer was passed through the chamber (step 5, Fig. 1).
No more than 5 L of buffer was passed through without regeneration of the Chelex matrix unless EDTA (≥1 mM) was added supplementary to Chelex treatment.
To measure thrombus formation along the whole capillary, lysis buffer was passed through each capillary and protein concentration of the lysates measured as an indication of platelet number.
The phosphate buffer was passed through Chelex 100 resin before it was used to prepare the solutions, and was used as a blank in the assay.
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Following recordings in the presence of hTpx2, a further 3 4 cell volumes of assay buffer were passed through the flow cell to make the final recordings following hTpx2 wash out.
Nuclei (in 1.5 mL buffer) were passed through a filter of pore size 40 μm, stained with 50 μg/mL propidium iodide (PI) and analyzed on a FACS Calibur flow cytometer (Becton, Dickinson and Company, USA) with CellQuest software.
Protein samples for ICP-MS analysis were digested with HNO3 and then refluxed for 30 min. All buffers were passed through a column of Chelex 100 (Bio-Rad) to remove trace metal contamination.
At the start of separation 100 0 Buffer A: Buffer B was passed through the column, and adjusted gradually to 0 100 Buffer A: Buffer B by the end of the analysis, which lasted 45 min. Buffer A consisted of 80% (v/v) sodium acetate (pH 5.9), 20% (v/v) methanol and 5 15 ml tetrahydrofuran/l and Buffer B consisted of 80% (v/v) methanol and 20% (v/v) sodium acetate (pH 5.9).
After the buffer solution was passed through for 6.5 h, HSA solution was passed through at 5 mL h-1 for 15 to 16 h.
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