IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/07/01/Shan Shen
From OpenWetWare
Jump to navigationJump to search
4:00 Proportion 3:1 is enough for the ligation and CIP is necessary for the double digestion.
Choose the plates of double digestion with CIAP to pick the single clones and each plate pick 5 clones. Incubate them at 37℃.
17:30 Mimiprep the plasmids of ligation
20:00 Electrophoresis to test the ligation products
The order and the amount of the samples: marker 5μL, plasmids of T7 promoter 2μL, ligation products (1-12M) 2μL, ligation products (1-12O) 2μL.
Results:
O3 is weird, others are correct.
21:00 Digestion to test the ligation products,Digestion System:20μL
| Plasmids | 1μL |
| EcoR1 | 1μL |
| Pst1 | 1μL |
| Buffer | 2μL |
| ddH2O | 15μL |
21:50 Start to digest.
