Team:UCLA/Notebook/Spider Silk Genetics/11 August 2015
Contents
08/11/2015
BsaI Digestion of M2AB, BC and CA
| 2AB | 2BC | 2CA | |
| Cutsmart | 5 uL | 5 uL | 5 uL |
| Template | 35 uL | 19.5 uL | 18.2 uL |
| ddH2O | 6 uL | 21.5 uL | 22.8 uL |
| BsaI | 4 uL | 4 uL | 4 uL |
50 C for 2hr 65 C for 20 min
ICA M1-15
Same as previous protocol
PCR amplified eluted ICA product
| Q5 buffer | 10 uL |
| F-03 Primer | 2.5 uL |
| G-03 Primer | 2.5 uL |
| Template | 1 uL |
| ddH2O | 32.5 uL |
| Q5 | 0.5 uL |
| 98C | 30 sec |
| 98C | 10 sec |
| 66C | 20 sec |
| 72C | 50 sec |
| x25 | |
| 72C | Hold |
| 12C | Hold |
Results
- Lane 1 and 2
- Brightest band was not the right size
- Band of expected size could barely be seen in lane 1 but not in lane 2
- Might have to redo ICA
- All BsaI digestion samples had the right band
T4 ligation of digested products from 08/10/2015
Transformed with ligation products