Team:UCLA/Notebook/Spider Silk Genetics/10 August 2015
08/10/2015
Redid PCR to amplify M1-12 ICA product
| 5X Q5 buff | 10 uL |
| 10mM dNTPs | 1 uL |
| F-03 | 2.5 uL |
| G-03 | 2.5 uL |
| Template | 1 uL |
| ddH2O | 32.5 uL |
| Q5 | 0.5 uL |
| 98C | 30 sec |
| 98C | 10 sec |
| 66C | 20 sec |
| 72C | 40 sec |
| x23 | |
| 72C | 2 min |
| 12 C | Hold |
- Changed extension time to 40 sec instead of 35 sec
- Used new primers from stock
Ran samples on 1% gel with 1kb ladder
Gel purified: 11ng/uL in 30uL
EcoRI+PstI Digestion of M1-12
| EcoRI | 1 uL |
| Pst I | 1 uL |
| 2.1 buffer | 5 uL |
| DNA | 33 uL |
37C for 1.5 hr 65C for 20 min