IGEM:Peking University/2008/Experiment/PKU 0807018 G1 1

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PKU_080718_G1_1


Enzyme cutting-pADH-lac2(PCR product)
System for double ligation
Volume (μL)
Promoter20
BamHI1
SacI1
0.5×K Buffer2.5
ddH2O25.5
System for single ligation
Volume (μL)
Promoter20
XhoI2
SacI2
1×M Buffer5
ddH2O21



'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)
Promoter20
BamHI1×3
SacI1
0.5×K Buffer2.5
ddH2O25.5
System for single ligation
Volume (μL)
Promoter20
XhoI2×3
SacI2
1×M Buffer5
ddH2O21



Single Ligation-pGREG504-pAID-lac2
Volume (μL)
lac2 4
pGREG504 1
ligation buffer5


Double Ligation-pGREG504-pAID-lac2-AID
Volume (μL)
lac2 3
AID 3
pGREG504 1
ligation buffer7


Control C1
Volume (μL)
lac24
ddH2O 4
ligation buffer5


Control C2
Volume (μL)
pGREG504 1
ddH2O 4
ligation buffer7


Control C3
Volume (μL)
lac23
ddH2O 4
ligation buffer7


Control C4
Volume (μL)
AID3
ddH2O 4
ligation buffer7


Control C5
Volume (μL)
AID3
lac23
ddH2O 1
ligation buffer7



Gel running-testing-pADH-lac1/S/tet
Gel running-testing-digestion-lac2/AID


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