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After aspiration and washing, 100 l of substrate solution was added to each well and incubated for 30 min at room temperature. Then, 100 L of a stop solution was added, and color changes were measured at 450 nm using a BioTek ELISA reader

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Injections are particularly appropriate for those with: Pernicious anaemia, an autoimmune condition affecting B12 absorption Conditions affecting the stomach or small intestine, such as Crohns disease or coeliac disease Those who have had gastric surgery or bariatric procedures Strict vegans and vegetarians not meeting their B12 needs through food or oral supplements For those who can absorb B12 normally, oral supplements are generally an effective and well-evidenced alternative to injections