Rna pico sensitivity assay

Fifteen FFPE sections of 4 μm thickness from each sample were stored in tubes prior to RNA extraction, as previously described [12 (link),13 (link)]. Briefly, total RNA was isolated from FFPE samples using the Quick-RNA FFPE Kit (Zymo Research, Irvine, CA, USA). Following a DNAse I digestion using the Baseline-ZERO kit (Epicentre, Illumina, San Diego, CA, USA), the RNA concentration was measured with the Qubit RNA HS Assay Kit (Life Technologies, Carlsbad, CA, USA) on a Qubit Fluorometer (Life Technologies, Carlsbad, CA, USA). The RNA quality was determined with the RNA Pico Sensitivity Assay (PerkinElmer, Waltham, MA, USA) on a LabChip GXII Touch (PerkinElmer, Waltham, MA, USA).

After melting the paraffin block, the pterygium, as well as the control FFPE samples were stored in tubes until RNA isolation, which was performed as previously described (21 (link), 22 (link)). Briefly, total RNA was isolated from FFPE samples using the Quick-RNA FFPE Kit (Zymo Research). Following a DNAse I digestion using the Baseline-ZERO kit (Epicentre), the RNA concentration was measured with the Qubit RNA HS Assay Kit on a Qubit Fluorometer (Life Technologies). The RNA quality was determined with the RNA Pico Sensitivity Assay on a LabChip GXII Touch (PerkinElmer).