IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Core/2009/07/21/Shan Shen
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00:40
Digest SupD and terminator plasmids.
System of SupD:
Total | 20μL |
Plasmids | 6μL |
EcoR1 | 1μL |
Spe1 | 1μL |
Buffer | 2μL |
ddH2O | 10μL |
System of terminator 1-23L:
Total 20μL |
Plasmids 6μL |
EcoR1 1μL |
Xba1 1μL |
Buffer 2μL |
ddH2O 10μL |
1:00
Start to digest.
10:00
Electrophoresis to test the digestion products.
The order and the amount of the samples: marker 5μL, SupD digestion, 1-23L plasmids, 1-23L digestion.
Results:
The digestions are successful.
11:10
Recycle the fragments of SupD.
13:10
Ligation of the vectors of 1-23L and inserts of SupD.
System of ligation: 10μL
Control system | 1:7 system | |
Vectors | 1μL | 1μL |
Inserts | 0μL | 7μL |
Buffer | 1μL | 1μL |
Ligase | 1μL | 1μL |
ddH2O | 7μL | 0μL |
13:10
Ligation starts.
14:21
Miniprep bi-stable 1,2,3,SupD1 and 1-9H.
Concentrations:
Number of the plasmids | Concentration(ng/μL) |
Bi-stable 1 | 160 |
Bi-stable 2 | 200 |
Bi-stable 3 | 250 |
SupD1 | 250 |
1-9H | 110 |
17:20
Transform the ligation products of SupD+1-23L backbone.