Table 1. Large genomic segments directly cloned from bacteria, mammalian cells and human blood with ExoCET.
| Target | Source | Genome (Mb) | Digestion enzymes | Fragment (kb) | Vector | c.f.u. (/ml) | Correct/checked |
|---|---|---|---|---|---|---|---|
| plu3535-3532 | P. luminescens DSM15139 | 5.69 | XbaI | 38 | pBAC2015 | 1815±132 | 12/12 |
| plu2670 | P. luminescens DSM15139 | 5.69 | XbaI+XmaI | 53 | p15A | 787±194 | 10/12 |
| salinomycin cluster | S. albus DSM41398 | 8.38 | EcoRV | 106 | pBeloBAC11 | 425±91 | 2/24 |
| salinomycin cluster | S. albus DSM41398 | 8.38 | Cas9 | 106 | pBeloBAC11 | 260±14 | 1/24 |
| Wnt4 | Mouse melanoma B16 cell | 2800.06 | SwaI | 45 | p15A | 76±16 | 8/25 |
| Lmbr1l-Tuba1a | Mouse melanoma B16 cell | 2800.06 | SwaI | 53 | p15A | 52±6 | 1/12 |
| Prkar1a | Mouse melanoma B16 cell | 2800.06 | HpaI | 8 | p15A | 205±17 | 10/12 |
| IGFLR1-ARHGAP33 | Human blood | 3221.49 | BstZ17I | 41 | p15A | 275±76 | 5/48 |
| ZBTB32-LIN37 | Human blood | 3221.49 | NdeI | 45 | p15A | 115±35 | 2/48 |
| Dpy30 | Mouse melanoma B16 cell | 2800.06 | BamHI+KpnI | 8.7 | p15A | 273±18 | 9/12 |
| DPY30 | HEK 293T cell | 3221.49 | SpeI | 9.1 | p15A | 40±10 | 17/24 |
| DPY30 | Human blood | 3221.49 | SpeI | 9.1 | p15A | 45±2 | 5/24 |
| Oct4-Venus | Mouse R1 ES cells | 2800.06 | EcoRV+PacI | 9.6 | p15A | 34±1 | 9/36 |
| Nanog-Cherry | Mouse R1 ES cells | 2800.06 | NdeI | 13 | p15A | 49±12 | 17/54 |
| Gata2-Venus | Mouse GM8 ES cells | 2800.06 | BstZ17l | 16.8 | p15A | 212±27 | 5/45 |
| Mll4 (1) | Mouse R1 ES cells | 2800.06 | SspI+SpeI | 17.1 | p15A | 127±38 | 7+3/24 |
| Mll4 (2) | 323±65 | 2+2/36 | |||||
| Mll4 (3) | 142±27 | 6+9/72 | |||||
| Mll4 (4) | 483±91 | 3+5/36 |
All experiments were done in triplicate; c.f.u. includes standard deviation and fidelity was monitored by restriction analysis of the indicated number of colonies. For the Mll4 experiments, fidelity shows the targeted allele + wt allele/colonies examined. DNA analyses are shown in Supplementary Figure S6.