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One milliliter of a PBS-based reaction mixture containing propidium iodide (20 g/mL), RNase (0.2 g/mL), and 0.1% Triton X-100 (Sigma) was added to the cell samples at a density of 1 10 6 cells/mL and incubated in the dark for 40 min before a DNA content analysis was conducted through flow cytometry

While the protective effect of folate against NTD is well established, only a few studies have investigated the role of other methyl group donors, including choline and betaine, in the occurrence of NTDs

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have a history of pancreatitis or thyroid cancer