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Components of PCR and thermal conditions.
| Item | Concentration | Amount | Temperature | Time | Cycle |
|---|---|---|---|---|---|
| GG template | 1/10 diluted | 1 μL | 94 °C | 2 min | 1 |
| Aa_g1g2_F2 | 10 μM | 2 μL | 94 °C | 15 s | 40 |
| Reverse primer * | 10 μM | 2 μL | 60 °C | 20 s | |
| 10X KOD Buffer | 5 μL | 68 °C | 1 min/kb ** | ||
| dNTP | 2 mM | 4 μL | 68 °C | 5 min | 1 |
| MgSO4 | 25 mM | 3 μL | 4 °C | Hold | 1 |
| KOD -Plus- | 1 U/μL | 1 μL | |||
| Distilled Water | to 50 μL |
* pBAtC_4sg reverse primer: Aa_g3g4_R2. * pBAtC_6sg reverse primer: Aa_g5g6_R2. ** 4sg time: 40 s; 6sg time: 1 min 10 s.
