| Component | Amount (per reaction, μL) | Final |
|---|---|---|
| BbsI-digested pX330 (20 ng/μL) | 2.5 | 50 ng |
| Annealed oligonucleotide duplex (1:250) | 1 | 4 nM |
| 2× Rapid ligation buffer | 5 | 1× |
| T4 DNA ligase | 1 | 3 U |
| ddH2O | 1.5 | - |
| Total volume | 11 | |
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