Silver: Ligation
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Use Roche's Rapid DNA Ligation Kit, stored at -20 °C.
Keep the total mass of DNA below 200 ng.
- Mix:
- Add 10 µL of freshly mixed tube #1 (2x T4 DNA ligase buffer) or 2uL of 10x T4 DNA ligase buffer
- Add 1 µL tube #3 (T4 DNA ligase).
- Mix well, incubate for 15 min. at room temperature.
- Proceed to transformation. Store excess ligation mixture at -20 °C.