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| (1) System: | 2×PrimeStar | 12.5μL |
| Forward primer | 1~2.5μL |
| Reverse primer | 1~2.5μL |
| Bacteria solution | 1~2μL |
| dd water | up to 25μLL |
| (2) Steps: | Step 1 | 94℃ |
| Step 2 | 94℃ |
| Step 3 | Annealing temperature |
| Step 4 | 72℃ |
| Step 5 | go to Step 2, 25~35 cycles |
| Step 6 | 72℃ |
| Step 4 | Pause at 4℃ |
| (1) System: | 2×PrimeStar | 12.5μL |
| Forward primer | 1μL |
| Reverse primer | 1μL |
| Template DNA | 0.5μL |
| dd water | up to 25μL |
| (2) Steps: | Step 1 | 94℃ |
| Step 2 | 94℃ |
| Step 3 | Annealing temperature |
| Step 4 | 72℃ |
| Step 5 | go to Step 2, 25~35 cycles |
| Step 6 | 72 ℃ |
| Step 7 | Pause at 4℃ |
| Buffer I: | 100mM RbCl | 1.2g/100mL |
| 45mM MnCl2·4H2O | 0.891g/100mL |
| 30mM KAc(pH 7.5) | 0.294g/100mL |
| 10mM CaCl2·2H2O | 0.147g/100mL |
| glycerol (pH 6.8) | 15% (w/v) |
| Buffer II: | 10mM MOPS | 0.209g/100mL |
| 10mM RbCl | 0.12g/100mL |
| 75mM CaCl2·2H2O | 1.1025g/100mL |
| glycerol (pH 6.8) | 15% (w/v) |
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