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To prepare membranes containing either FLVCR2 or PNS1 protein, 300 g of cells was thawed and resuspended in 750 ml lysis buffer (600 mM NaCl, 100 mM TrisHCl, pH 7.5), supplemented with 1 mM PMSF, followed by 5 1 min cycles of bead beating with 0.5-mm glass beads (BioSpec Products)

Although one genetic variant (PEMT rs7946) appeared more frequently among NTD cases, the takeaway was clear: choline levels alone dont strongly predict NTD risk

Immunol Rev (2019) 288:16177

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The preparation must be inserted strictly during the final phase of the formulation (cooling phase), at temperatures below 40C