PKU_080718_G1_1
- Enzyme cutting-pADH-lac2(PCR product)
- System for double ligation
|
Volume (μL)
|
| Promoter |
20
|
| BamHI |
1
|
| SacI |
1
|
| 0.5×K Buffer |
2.5
|
| ddH2O |
25.5
|
- System for single ligation
|
Volume (μL)
|
| Promoter |
20
|
| XhoI |
2
|
| SacI |
2
|
| 1×M Buffer |
5
|
| ddH2O |
21
|
- 'PCR- pGREG504-pADH-(lac1/lacS/tet)-50ul
- Using program lac12MQT
- Gel running and extraction-promoter(PCR product)
- Enzyme cutting-AID
- lanes: intact AID(control)|λmarker|Promoter(single)|promoter(double)|AID|AID
- System for double ligation
|
Volume (μL)
|
| Promoter |
20
|
| BamHI |
1 |
×3
|
| SacI |
1
|
| 0.5×K Buffer |
2.5
|
| ddH2O |
25.5
|
- System for single ligation
|
Volume (μL)
|
| Promoter |
20
|
| XhoI |
2 |
×3
|
| SacI |
2
|
| 1×M Buffer |
5
|
| ddH2O |
21
|
- Single Ligation-pGREG504-pAID-lac2
|
Volume (μL)
|
| lac2 |
4
|
| pGREG504 |
1
|
| ligation buffer |
5
|
- Double Ligation-pGREG504-pAID-lac2-AID
|
Volume (μL)
|
| lac2 |
3
|
| AID |
3
|
| pGREG504 |
1
|
| ligation buffer |
7
|
- Gel running-testing-pADH-lac1/S/tet
- Gel running-testing-digestion-lac2/AID
|
Group 1
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