IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/06/30/Zhangs
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Double digestion of CI
System for front insert:
| 10μL | plasmid |
| 2μL | 10×H beffer |
| 1.5μL | EcoRI |
| 1.5μL | SpeI |
| 5μL | ddH2O |
| 20μL | Total |
4 h digested
Electrophoresis
Result (from left to right)
CI, CI plasmid control, marker
Gel: 0.072gPN volume: 216μL
Gel extraction for CI insert
Ligation Insert:CI Vector:1-23L, 1-4H
System
(1)7:1 system
| 7μL | CI insert |
| 1μL | vector |
| 1μL | 10× Ligase buffer |
| 1μL | Ligase |
| 10μL | Total |
(2)3:1 system
| 3μL | CI insert |
| 1μL | vector |
| 1μL | 10× Ligase buffer |
| 1μL | Ligase |
| 4μL | ddH2O |
| 10μL | Total |
Control
| 0 | CI insert |
| 1μL | vector |
| 1μL | 10× Ligase buffer |
| 1μL | Ligase |
| 7μL | ddH2O |
| 10μL | Total |
6 samples in total, 2 for 1-23L, 2 for 1-4H, 2 for 1-23L & 1-4H controls
