GSM4971904: Mouse_CNV_S4; Mus musculus; RNA-Seq
RNA isolation from FFPE specimens was carried out as previously described. Briefly, total RNA was extracted from FFPE samples using the Quick-RNA FFPE Kit (Zymo Research, Irvine, California). Following DNAse I digestion using the Baseline-ZERO Kit (Epicentre, Madison, Wisconsin), the RNA concentration was quantified using the Qubit RNA HS Assay Kit on a Qubit Fluorometer (Life Technologies, Carlsbad, California). RNA quality was determined via the RNA Pico Sensitivity Assay on a LabChip GXII Touch (PerkinElmer, Waltham, Massachusetts). RNA sequencing was performed using massive analysis of cDNA ends (MACE), a 3' RNA sequencing method, as previously described. We recently demonstrated that MACE allows sequencing of FFPE samples with high accuracy. Briefly, 20 barcoded libraries comprising unique molecule identifiers (six mouse CNV mem-branes, six mouse RPE-choroidal control samples, four human CNV membranes and four human RPE-choroidal control tissues) were sequenced on the NextSeq 500 (Illumina) with 1 × 75 bp. PCR bias was removed using unique molecular iden-tifier.