| Component | Volume | Final Concentration |
|---|---|---|
| 10× PCR Buffer for KOD -Plus- Neo | 5μl | 1x |
| 2 mM dNTPs | 5μl | 0.2 mM each |
| sgRNA IVT F (10μM) | 2μl | 0.4μM |
| sgRNA IVT R (10μM) | 2μl | 0.4μM |
| KOD -Plus- Neo (1.0 U/μl) | 1μl | 1.0 U/50 μl |
| px330(100ng/μl) | 0.5μl | 1ng/μl |
| 25 mM MgSO4 | 3μl | 1.5 mM |
| ddH2O | 31.5μl | |
| Total | 50μl |
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