Table 2.
Protection of transposase-loaded DNA from restriction.
| Experiment | Mycoplasma strain | pH of culture at harvest | Number of mycoplasma colonies after transformation | ||
|---|---|---|---|---|---|
| Tn5 designer transposome (puromycin) | Plasmid | No DNA | |||
| 1 | JCVI-syn1.0 | 6.6 | 117,000 | 7350 | 0 |
| JCVI-syn1.0 Δ1-6 | 6.8 | 222,450 | 696,000 | 0 | |
| 2 | JCVI-syn1.0 | 6.7 | 66,960 | 3660 | 0 |
| JCVI-syn1.0 Δ1-6 | 6.7 | 106,980 | 223,980 | 0 | |
Four hundred nanograms of DNA were used for transposome samples. Four hundred and one hundred nanograms were used in experiments 1 and 2, respectively, for plasmid samples. Colony numbers normalized to represent the whole transformation mixtures are shown. The pH values of the cultures used for transformation procedures are shown. pH of the medium decreases as mycoplasma cells grow, and the pH range of 6.6–6.8 is roughly associated with late log phase.