title: GSM4832554: GZS34 het; Mus musculus; RNA-Seq
library_construction_protocol: RNA from microdissected kidneys was immediately extracted by Tryzol, using the miRNA mini kit Qiagen for the extraction of total and miRNAs. The quality of the RNA samples was assessed on the Agilent Bioanalyzer system using the Agilent RNA 6000 Nano Kit (Agilent Technologies) and RNA with a Ring higher than 8,5-9 was used. 1-2 μg of total RNA were used for mRNA isolation using the Dynabeads® mRNA DIRECT™ Micro Kit (ThermoFisher Scientific). mRNA was digested with RNase III, purified, hybridized and ligated to Ion Adaptors, reverse transcribed, barcoded and amplified, using the Ion Total RNA‐Seq Kit v2 (ThermoFisher Scientific). The pools were then processed on an OneTouch 2 instrument and enriched on a One Touch ES station. Templating was performed using the Ion PI™ Template OT2 200 Kit (ThermoFisher Scientific) and sequencing with the Ion PI™Sequencing 200 Kit on Ion Proton PI™ chips (ThermoFisher Scientific) according to commercial protocols. RNA sequencing from E17.5 embryo kidneys (SAMPLES 7 to 10) was performed by FASTERIS-Genome Analyzer Service (Switzerland). High-throughput DNA sequencing using Illumina technology consisted in processing in vitro samples to generate a library of short inserts (the DNA Colonies Template Library). The library was sequenced on the Illumina HiSeq 2000. For each lane 130-150 million DNA colonies producing pass filter sequences were assured. The read lengths are 1x50 or 1x100 base pairs (bp) for single-reads runs using the forward sequencing primer. The inserts can also be sequenced from both ends using a 'forward' and a 'reverse' sequencing primer, generating pairedreads of 2x100 bp. The data were processed using bioinformatics tools to extract biologically useful information.