IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/07/08/Zhangs
From OpenWetWare
Transformation for the ligation product: T7p+2M/1H/11N/2G/2I/5N/1J RBS+CI+1-23L, 7 samples in total.
Double digestion for 1-2K+CI+1-23L insert again (yesterday failed)
| 1.5μL | XbaI |
| 1.5μL | PstI |
| 2μL | 10×M buffer |
| 10μL | 1-2K③ plasmid |
| 5μL | ddH2O |
| 20μL Total |
Electrophoresis and Gel Extraction for 1-2K+CI+1-23L and CI+1-23L insert (for 1-5J vector)
| 2K+CI+1-23L | 0.081g | 243μL |
| CI+1-23L for 5J | 0.100g | 300μL |
Ligate the 1-2K+CI+1-23L insert and T7 promoter vector
System
| 3μL | 1-2K+CI+1-23L insert |
| 1μL | T7p vector |
| 1μL | 10× Ligase buffer |
| 1μL | Ligase |
| 4μL | ddH2O |
| 10μL | Total |
Ligate the CI+1-23L insert and 1-5J RBS vector
System
| 3μL | CI+1-23L insert |
| 1μL | 1-5J vector |
| 1μL | 10× Ligase buffer |
| 1μL | Ligase |
| 4μL | ddH2O |
| 10μL | Total |
