IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/06/28/Shan Shen
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14:00 Miniprep the plasmids of T7 promoter, 1-12M, 1-12O.
The concentration of the plasmids:
| T7 promoter | 305ng/μL |
| 1-12M | 280ng/μL |
| 1-12O | 295ng/μL |
15:00 Digest the plasmids of T7 promoter, 1-12M and 1-12O
Digestion System for plasmids of T7 promoter: 20μL
| Plasmids | 3μL |
| Pst1 | 1μL |
| Buffer 1*H | 2μL |
| dd H2O | 14μL |
Digestion System for plasmids of 1-12M and 1-12O: 20μL
| Plasmids | 4μL |
| Xba1 | 1μL |
| Pst1 | 1μL |
| Buffer 1*M | 2μL |
| dd H2O | 12μL |
17:10 Start to digest.
22:00 Double digestion of plasmid of T7 promoter.
Digestion System for plasmids of T7 promoter: 20μL
| Plasmids | 3μL |
| Xba1 | 1.5μL |
| Pst1 | 1.5μL |
| Buffer 1*M | 2μL |
| dd H2O | 12μL |
23:25 Electrophoresis to test the digestion efficiency (17:10).
The order and the amount of the samples: marker 10μL, plasmids of T7 promoter 10μL, digestion products of T7 promoter 10μL, plasmids of 1-12M 10μL; digestion products of 1-12M, plasmids of 1-12O, digestion products of 1-12O.
Results:
The plasmids have been cut downright and are suitable for recycling.
