Table 3.
Hierarchical assembly of a 218 kb DNA fragment using a BsaI-based MetClo system
| Plasmid | Vector | Inserts | Size | Success ratea | ||||||
|---|---|---|---|---|---|---|---|---|---|---|
| pMXBG_3A1 | pMXBG_A7I | pMOLK_2A1 | pMOLK_2B1 | pMOLK_2C1 | pMOLK_2D1 | pMOLK_2E1 | pMOLK_2F1 | pMOLK_2G1 | 54 kb | 75% (3/4) |
| pMXBG_3I1 | pMXBG_I7G | pMOLK_2I1 | pMOLK_2A2 | pMOLK_2B2 | pMOLK_2C2 | pMOLK_2D2 | pMOBK_2E2 | pMOLK_2F2 | 55 kb | 50% (2/4) |
| pMXBG_3G1 | pMXBG_G7F | pMOLK_2G2 | pMOLK_2I2 | pMOLK_2A3 | pMOLK_2B3 | pMOLK_2C3 | pMOBK_2D3 | pMOLK_2E3 | 54 kb | 50% (2/4) |
| pMXBG_3F1 | pMXBG_F7E | pMOLK_2F3 | pMOLK_2G3 | pMOLK_2I3 | pMOLK_2A4 | pMOLK_2B4 | pMOLK_2C4 | pMOLK_2D4 | 54 kb | 50% (2/4) |
| pMXBK_4A1 | pMXBK_A7E | pMXBG_3A1 | pMXBG_3I1 | pMXBG_3G1 | pMXBG_3F1 | 218 kb | 20%b |
aCalculated based on the white colonies selected following blue-white screening.
b14/27 verified by PCR screening, of which 3/8 verified by restriction digest.