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How do I prepare a firefly luciferase standard curve? answer
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LabFact #32
Vary the incubation time and temperature for successful ligation. Blunt-end ligations benefit from lower temperatures (4–16°C) and longer incubation times (4 hours to overnight). Cohesive-end ligations can generally be performed at higher temperatures (15–20°C) with shorter incubation times (3–16 hours).
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