Table 1.
Comparison of the T‐DNA copy numbers estimated by Southern blot, qPCR, TAIL‐PCR and ddPCR for eight plants representing three generation (T0, T1 and T2) and corresponding control wild type (WT) of N. tabaccum. For TAIL‐PCR the T‐DNA copy number was assessed with the use of three different reaction sets with primers differing in degeneration: AD1, AD2 and AD3, with 64‐, 128‐ and 256‐fold degeneracy, respectively. For qPCR and ddPCR the T‐DNA copy number was derived from the estimated sample concentration of T‐DNA relative to actin (± se; n = 4 for qPCR; n = 2 for ddPCR).
| T‐DNA copy number | ||||||
| Southern blot | qPCR | TAIL‐PCR | ddPCR | |||
| Plant ID | AD1 | AD2 | AD3 | |||
| VPZ‐23 T0 | 2 | 2.37 ± 0.06 | 1 | 1 | 2 | 1.83 ± 0.11 |
| VPZ‐23 T1.3 | 2 | 2.17 ± 0.18 | 1 | 1 | 2 | 1.90 ± 0.20 |
| VPZ‐23 T1.4 | 0 | 0 ± 0.00 | 0 | 0 | 0 | 0 ± 0.00 |
| VPZ‐23 T1.5 | 4 | 4.83 ± 0.19 | 1 | 1 | 2 | 3.74 ± 0.04 |
| VPZ‐23 T1.6 | 4 | 5.37 ± 0.15 | 1 | 1 | 2 | 4.21 ± 0.28 |
| VPZ‐23 T1.8 | 2 | 2.29 ± 0.62 | 0 | 1 | 1 | 1.87 ± 0.04 |
| VPZ‐23 T2.2 | 4 | 4.40 ± 0.12 | 0 | 0 | 1 | 4.24 ± 0.11 |
| VPZ‐23 T2.3 | 4 | 4.72 ± 0.16 | 1 | 1 | 2 | 4.18 ± 0.16 |
| WT | 0 | 0 ± 0.00 | 0 | 0 | 0 | 0 ± 0.00 |