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Push the plunger forward slowly until the bottom of the plunger seal aligns exactly with your target dose marking

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Subsequently, 180 l fresh culture medium and 20 l of samples (0.05858000 M in sterile water) were added per well and incubated for 24, 48 and 72 hours

This has nothing to do with b12 status and everything to do with excessive caloric deficit and insufficient protein intake relative to the rate of weight change