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Table 2 Measurement of viral RNA in concentrated lentiviral vector preparations

From: Comparison of lentiviral vector titration methods

  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    
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
  1. 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.