Table 14.
PCR components and thermal conditions.
| Item | Concentration | Amount | Temperature | Time | Cycle |
|---|---|---|---|---|---|
| Colony | (5 ng) | 95 °C | 5 min | 1 | |
| U6-26P_F | 10 μM | 1 μL | 95 °C | 30 s | 30 |
| NosP_Rev * | 10 μM | 1 μL | 55 °C | 30 s | |
| 10X Buffer | 2 μL | 72 °C | See footer ** | ||
| dNTP | 2.5 mM | 1.6 μL | 72 °C | 5 min | 1 |
| DNA polymerase | 5 U/μL | 0.1 μL | 8 °C | Hold | 1 |
| Distilled Water | to 20 μL |
* NosP_Rev sequence: AAGTCGCCTAAGGTCACT. ** pBAtC_4sg, expected PCR band size: 1159 bp. ** pBAtC_6sg, expected PCR band size: 1513 bp. ** Use different elongation times based on the DNA-polymerase performance.