FAQ: Why isn't SmaI cutting?

1. SmaI needs K+ to work. Use in NEBuffer 4, or supplement with KCl.
2. SmaI is blocked by CpG methylation.
3. The reaction temperature for SmaI is 25°C. Incubation at 37°C gives 50% activity. SmaI has a half-life of 15 minutes at 37°C.

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