Figure 4.

Impact of different staining and labeling protocols on the amount of EF-1a mRNA detected in cryo-sections of rabbit appendix using quantitative real-time RT-PCR. All methods that comprise a single incubation of only 5 minutes (bars 2 to 5) yielded comparable mRNA recovery and did not significantly differ from untreated controls. In contrast, an incubation of 60 minutes led to a significant loss of detectable EF-1a copies (P < 0.05; asterisk). The difference of 9.3 Ct units, which equals a factor of more than 600, between the 5-minute and the 60-minute incubations corresponds to a quantitative loss of 99.8% EF-1a mRNA. Bars represent means and SD of six samples per group.