Table 2 Comparison of time investments to complete traditional cloning versus Pyrite cloning
From: Pyrite cloning: a single tube and programmed reaction cloning with restriction enzymes
Step | Traditional cloning | Time investment | Pyrite cloning | Time investment |
|---|---|---|---|---|
1a | Digest vector | 1–24 h | Prepare and incubate reaction | 18–25 h |
1b | Digest insert(s) | 1–24 h | ||
2 | Gel electrophoresis | 15–45 min | Transform | 1–2 h |
3 | Gel purification | 1–2 h | ||
4 | Ligation reaction | 16 h | ||
5 | Deactivate ligase | 10 min | ||
6 | Transform | 1–2 h | ||
Total steps | 6 | 2 | ||
Total labor time | 2–4 h | 0.5–1 h | ||
Total incubation time | 19–43 h | 19–26 h |