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Time consideration of the key procedures
| Procedure | Time (h) |
|---|---|
| Cytosol preparation | 6 |
| Total membrane preparation | 2 |
| Lipidation reaction | 1.5 |
| LC3 protein purification | 24 |
| Three-step membrane fractionation | 24 |
| Membrane recruitment | 1.5 + 1.5 (reaction + centrifugation) |
| Small autophagosome precursor formation | 1.5 + 1.5 lipidation |
