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After this time, the unused portion should be discarded

doi:10.1111/j.1365-2133.2009.09216.x

Neutrophils were pelleted at 250 g for 6 min, washed in RPMI 1640 with 2% FBS, 2 mM L-glutamine and 10 mM HEPES, and centrifuged at 250 g for 4 min

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This means you may experience symptoms such as: Bloating Constipation Diarrhea Gas Nausea However, its important to keep in mind that gastrointestinal issues alone may not signal a vitamin B12 deficiency