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Cells were harvested, washed once with phosphate-buffered saline (PBS), and prepared for cell cycle analysis using the "Cycle Test Plus Kit" (BD, San Jose, USA).
The cells used for the cell-cycle analysis were stained with propidium oxide (100 μg/mL) using the Cycle Test Plus DNA Reagent Kit (BD Biosciences) and were analyzed by flow cytometry (FACScan; BD Biosciences) using an instrument equipped with the CellQuest software program (BD Biosciences).
HT-29 cells were prepared and treated as described, and then the percentage of cells in each phase of the cell cycle was analyzed by flow-cytometry, using the Cycle TEST PLUS DNA Reagent Kit (BD Pharmingen, San Jose, CA, USA), which is based on the measurement of the DNA content of nuclei labeled with propidium iodide (PI), according to manufacturer's instructions.
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Continue using the cycle lane.
Menstrual cycle dating was determined using the cycle history.
Therefore, the cell cycle status of the cells was studied by FACS analysis using the Cell Cycle Test (Becton Dickinson).
Since both tetrapeptide could induce an increase of cells in G0/1 phase with the Cycle Test™ kit was used because it was easier than Brdu incorporation to evaluate few modification in cell cycle repartition.
After 16 h, the cells were harvested and stained with propidium iodide using Cycle Test plus (BD Biosciences) according to the manufacturer's protocol.
Thus, this study reports the inhibition effect of glycine, alanine, proline, serine and arginine on the carbon dioxide hydrate phase equilibrium condition using the isochoric T-cycle method in the pressure range of 2.53 4.0 MPa.
Following amplification, levels of mRNA expression for the selected gene sequences were normalized relative to either GAPDH or β-actin expression using the comparative cycle time (C t ) method [ 41, 42].
Differences in the number of cycles of further chemotherapy/systemic therapy will be tested using the Student's t-test.
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