Stock ECE112, 166, 153
- Plasmid miniprep (2 tubes of ECE112, 2 tubes of ECE166 and 1 tube of ECE153)
- Nanodrop
Transformation of Bacillus
- For strain IA751 : transform with ECE166, 171, 153 and 112
- For strain IA771 : transform with ECE166, 171
- for glycerol stocks (one with 100μL of medium B and glycerol, and one with only glycerol) : transform with ECE166
- We used the same protocol than before with a few changes :
- add sterile 50% glucose in medium A
- don't keep samples after using them for spectrophotometer
- spin each tubes and keep only 100μL of medium A (to concentrate cells) before adding DNA
Time (min) |
0 |
20 |
40 |
60 |
80 |
100 |
120 |
130 |
140
|
OD650 for IA751 |
0.1817 |
0.1870 |
0.1929 |
0.2115 |
0.2543 |
0.3291 |
0.3695 |
0.4188 |
0.4224
|
OD650 for IA751 |
0.1818 |
0.1874 |
0.1907 |
0.2088 |
0.2452 |
0.3182 |
0.3575 |
0.4228 |
0.4314
|
- For glycerol stocks, resuspend cells into 100μL of medium B, add DNA and wait for 30min, end plate them
|