Confirmation of polysome profiling with RT-PCR. In an independent P. falciparum infection experiment, RNA samples collected from individual sucrose gradient fractions were not pooled together. RT-PCR was performed to display the distribution of mRNA in each fraction. Due to the presence of heparin in the extraction buffer, all RNA samples were treated with heparinase I (Sigma H2519) for 4 hours prior to RT-PCR in a protocol based on Izraeli S, et al. . INF: P. falciparum-challenged mosquitoes. UNI: control mosquitoes. DAP: spike-in RNA control.
Mead et al. BMC Genomics 2012 13:366 doi:10.1186/1471-2164-13-366