The Process of Plasmid Preparation
| A. Cultivation | ||||
| Process | Person | Tools | Vacuum pressure (inch Hg) | Time |
| Medium dispensing | 1 | HT medium | 1.75 | |
| dispenser | min/plate | |||
| Inoculation | 1 | 384 microtiter plate | 3 min/plate | |
| 96 pins | ||||
| 70% ethanol | ||||
| Cultivation | 1 | shaker (275 rpm) | 17 hr | |
| (for setting) | platform (30° angle) | |||
| 37°C room | ||||
| Culture transfer | 2 | 96-channel dispenser | 0.5 min/plate | |
| Vacuum filtration | 4 | vacuum manifold | 10 | 3 min/plate |
| vacuum pump (160 Torr) |
| B. Plasmid preparation | |||||
| Process | Stage | Solution | Volume (μl) | Vacuum pressure (inch Hg) | Time (min) |
| Lysozyme treatment | 2 | 1 | 25 | 2.5 | |
| Alkaline lysis | 3 | 2 | 50 | 2.5 | |
| Neutralization | 4 | neutralization | 200 | 2.5 | |
| Filtration | 5, 6, 7, 8 | 16 | 10 | ||
| Washing | 9,10 | washing | 600 | 16 | 5 |
| 11, 12 | 600 | 16 | 5 | ||
| Salt washing | 13, 14 | 80% ethanol | 300 | 20 | 5 |
| Drying | 15, 16 | 20 | 15 | ||
| 17, 18 | |||||
| 19, 20 |
| C. Elution | |||
| Process | Person | Tools | Time |
| Eluant dispensing | 1 | 96-channel dispenser | 0.5 min/plate |
| Centrifugation | 2 | centrifuge (1000g, for microtiter plate) | 10 min/16 |
| microtiter plate | plates | ||
| Drying | 1 | 60°C drying oven | overnight |