| Reagent | Amount for 1 well (nl) | Amount for 1 plate (μl) | Final concentration in mix |
|---|---|---|---|
| Nuclease-free water | 1,347.5 | 569.3 | |
| 5x Second strand buffer | 437.5 | 184.8 | 1.1 x |
| dNTP mix (10 mM) | 43.7 | 18.5 | 0.22 mM each |
| E. coli ligase (10 U/μl) | 15.7 | 6.6 | 0.08 U/μl |
| DNA polymerase I (10 U/μl) | 61.2 | 25.9 | 3.18 U/μl |
| Ribonuclease H (2 U/μl) | 15.7 | 6.6 | 0.016 U/μl |
| Total volume | 1,920.8 | 811.8 |
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