Table 1.
Changes made between protocols: T25 manual, T175 manual and T175 automated.
| Incubation time (min) | incubator temperature (°C) | Plasticware used to wash clumps | Time for colonies to settle down by gravity after washing (min) | Plasticware used to break clumps | Number of pipetting movements to break clumps | Split ratio | |
|---|---|---|---|---|---|---|
| T25 manual | 30 | 37.5 | Conical tube | 5 | 1000 μl tips | 5 | 1:10 |
| T175 manual | 35 | 37.5 | T175 flask | 10 | 10 ml pipette | 5 | 1:10 |
| T175 automated | 35 | 36.5 | T175 flask | 10–15 | 10 ml pipette | 7 | 1:7–1:5 |