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These factors contributed to the technique needing to be more sensitive than other assays

For NAD + and NADH assays, 20 L (for NAD + assay) or 30 L (for NADH assay) of the neutralized sample extract was combined with 90 L of assay mix consisting of 50 L of 0.1 M 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) with 2 mM EDTA (pH 7.5), 20 L of 1.2 mM 2,6-dichlorophenolindophenol, 10 L of 20 mM phenazine methosulfate, and 10 L of 2,500 units mL 1 aldolase dehydrogenase

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Benzodiazepines can cause sleep issues to go in the other direction as well, with some patients reporting paradoxical sleep/wakefulness issues, excessive sleep or exhaustion as part of their withdrawal process