Relationship between subjective assessment of chromatograms for total RNA degradation and quantification of FMR1 and DNMT1 mRNA, using real-time PCR. RNA samples from 5 lymphoblast cell lines were degraded at room temperature for 0 hr (A, E, I, M, Q), 18 hrs (B, F, J, N, R), 24 hrs (C, G, K, O, S), and 96 hrs (D, H, L, P, T), and assessed for total RNA integrity using Experion capillary electrophoresis ststem. Panels I-IV – RNA samples 496, 497, 488, 584, and 585, respectively. (◆) defines the boundary of each of the regions (see Figure 1). FMR1ex3/4, FMR1ex13/14 and DNMT1 mRNA arbitrary quantities were determined using real-time PCR relative standard curve method. Samples with the coefficient of variance greater than 30% between the duplicate reactions were omitted from the analysis.
Godler et al. BMC Clinical Pathology 2009 9:5 doi:10.1186/1472-6890-9-5