August, 22nd
Streak of E17-2, E18-2, J101-31, J101-E5 and ENTERO-4C5 on LB agar + Cm12.5 plate to test Plux promoter with different RBSs.
E24-1, E25-2, E26-1, E27-2 and E41N-1 purified DNA samples were sent to BMR Genomics for sequencing.
250 ml of M9 glycerol supplemented minimal medium were prepared.
Inoculum of T9002-ENTERO and ENTERO-RBS to test supernatants collected on August, 20th to test AiiA enzyme efficiency.
August, 23rd
T9002-ENTERO and ENTERO-RBS were diluted 1:500 in a final volume of 1 ml of M9 + Cm12.5.
Three colonies of E17-2, E18-2, J101-31, J101-E5 and ENTERO-4C5 were picked and inoculated in 1 ml of M9 + Cm12.5.
E19-2, E 20-2, J101-E7, J101-4C5 and ENTEOR-4C5 were streaked on a LB agar + Cm12.5 plate.
Unfortunately TECAN test could not be performed as the instrument did not work!
31 LB agar + Cm34 plates were prepared.
Last week sequencing were ready: E28-1 has not the desired sequence, while E37-2, E38-1, E39-1, E40-2 and E42-1 were correct.
Ptet-RBS30-luxI ligation was done again from E36 and E2:
| Ligation Name |
Vector |
Vector volume (μl) |
Insert |
Insert volume (μl) |
Buffer (μl) |
T4 Ligase (μl) |
| E43 |
E36 (S-P) |
3.5 |
E2 (X-P) |
4.5 |
1 |
1 |
"
