| Component | Amount (μl) Ligation | Amount (μl) Mock ligation | Final |
|
| |||
| Annealed oLigonucLeotides (50 μM) | 5.5 | 27.5 μM | |
| Digested and CIP-treated pLasmid | 1.5 | 1.5 | |
| NucLease-free H20 | 5.5 | ||
| T4 DNA Ligase buffer (5×) | 2 | 2 | 1× |
| T4 DNA Ligase (5 U μL−1) | 1 | 1 | 0.5 U |
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