| Ct value |
---|
DNA Standard | LTR primers | eGFP primers | WPRE primers |
---|
5.0 × 108 | 15.51 ± 0.01 | 15.65 ± 0.04 | 15.45 ± 0.06 |
5.0 × 107 | 17.57 ± 0.01 | 17.79 ± 0.10 | 17.51 ± 0.01 |
5.0 × 106 | 18.97 ± 0.06 | 19.11 ± 0.08 | 20.11 ± 0.05 |
5.0 × 105 | 22.06 ± 0.17 | 21.82 ± 0.07 | 22.59 ± 0.03 |
5.0 × 104 | 25.91 ± 0.05 | 24.06 ± 0.06 | 26.13 ± 0.04 |
RNA extracts
| Â | Â | Â |
CH-eGFP-WS
| 22.87 ± 0.05 | 22.5 ± 0.02 | 22.04 ± 0.03 |
CH-eGFP-WS
| 16.15 ± 0.02 | 16.57 ± 0.03 | 16.43 ± 0.05 |
- Primer/probe sets annealing to the front (LTR), the centre (GFP) or at the end (WPRE) of the genomic RNA of lentiviral vectors were used to determine to what extent full-length genomic vector RNA is incorporated into lentiviral vector particles. In one-step RT-qPCR assays with the different primer/probe sets comparable threshold cycles (Ct) were detected when amplifying a dilution series of the pCH-eGFP-WS transfer plasmid (DNA standard) or RNA extracts of two representative CH-eGFP-WS vector preparations. Mean Ct values ± standard deviation for 3 amplifications of the same sample are shown.