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The stirring process continued for another 12 h at 50 C, after which the mixture was introduced into 10 mL of deionized water while being sonicated

For days 48 of differentiation, cells were maintained in growth medium supplemented with 5 g ml 1 insulin until collection

Three cycles of such refinement were performed before inspecting the agreement between the atomic coordinates and electron density map in Coot 79 and building solvent molecules or adjusting side-chain positions to satisfy disagreements revealed in the difference Fourier map 79

AMB Express 11, 116113