Kidney samples were obtained from routine surgical nephrectomies and from leftover portions of diagnostic kidney biopsies. Only normal, non-neoplasmatic part of the tissue was used for further investigation. RNA sequencing was carried out on microdissected kidney tubules. Total RNA was isolated using the miRNeasy Kit (Qiagen) according to the manufacturer’s protocol. An additional DNase1 digestion step was performed to ensure that the samples were not contaminated with genomic DNA. RNA purity was assessed using the Agilent 2100 Bioanalyzer. Each RNA sample had an A260:A280 ratio above 1.8, a RIN>9, and an A260:A230 ratio above 2.2. Briefly, total RNA was converted to cDNA using oligodT, which was then used for Illumina sequencing library preparation. Single end 100 bp RNA-seq was carried on an Illumina HiSeq2500.
GEO Accession
GSM1465293