IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/07/03/Zhangs
Add CIAP into 12 RBS digesting vector samples.
Identify the digested ligation product of CI+1-23L/1-4H and RBS vector by electrophoresis
System:
| 1μL | sample |
| 1μL | DNA Dye |
| 4μL | ddH2O |
Marker:
| 5μL | Marker |
| 1μL | DNA Dye |
Digestions for RBS vectors failed and we digested again.
Samples: 1-11N, 1-1H, 1004-4C, 1-2M, 1-2I, 1-5N, 1-2G, 1-2K, 1-5J, 1004-3C, 1004-2G, 1-1J
System
| 6μL | RBS plasmid |
| 1.5μL | SpeI |
| 1.5μL | PstI |
| 2μL | 10×H buffer |
| 9μL | ddH2O |
| 10μL | Total |
Gel Extraction for CI+1-23L/1-4H insert
Gel: CI+1-23L 0.139gPN volume: 417μL
CI+1-4H 0.071gPN volume:213μL
Identify the RBS digestion products by electrophoresis
Results (from left to right)
Plasmid control, 1-2I, 1004-2G, 1004-4C, 1-5N, 1004-3C, 1-2K, 1-1H, 1-1J, 1-2G, 1-11N, 1-2M, 1-5J
There are no strips for thee 2008 iGEM parts 1004-2G, 1004-4C, 1004-3C. Perhaps the quality of the 2008 parts is bad.
DNA purification to extract RBS vector
Ligate the RBS vector and CI+Terminator insert
System
| 3μL | insert |
| 1μL | vector |
| 1μL | 10× Ligase buffer |
| 1μL | Ligase |
| 4μL | ddH2O |
| 10μL | Total |
