IGEM:Peking University/2008/Experiment/PKU 0807018 G1 1: Difference between revisions
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::'''Control | ::'''Control C5''' | ||
{|border="1" cellspacing="0" align="center" | {|border="1" cellspacing="0" align="center" |
Latest revision as of 04:05, 19 July 2008
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
- Enzyme cutting-pADH-lac1/lacS/tet
- 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 |
- Control C1
Volume (μL) | |
lac2 | 4 |
ddH2O | 4 |
ligation buffer | 5 |
- Control C2
Volume (μL) | |
pGREG504 | 1 |
ddH2O | 4 |
ligation buffer | 7 |
- Control C3
Volume (μL) | |
lac2 | 3 |
ddH2O | 4 |
ligation buffer | 7 |
- Control C4
Volume (μL) | |
AID | 3 |
ddH2O | 4 |
ligation buffer | 7 |
- Control C5
Volume (μL) | |
AID | 3 |
lac2 | 3 |
ddH2O | 1 |
ligation buffer | 7 |
- Gel running-testing-pADH-lac1/S/tet
- Gel running-testing-digestion-lac2/AID
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