title: GSM3317087: WT Brain Rep3 miRNA-seq; Drosophila melanogaster; miRNA-Seq
library_construction_protocol: RNA-seq: Total RNA was extracted by Trizol (ThermoFisher) following standard protocols. miRNA-seq: Total RNA including miRNAs was extracted using the Qiagen miRNeasy Mini kit following the manufacturers protocol. Small RNA was enriched from 2 µg total RNA by gel electrophoresis using a Bio-Rad PROTEAN® II xi Cell with a 15 % polyacrylamide/urea gel in 0.5 x TBE buffer. Samples were mixed with loading dye (2x; 89.75 % Formaldehyde, 10 % TBE (5x), 0.05 % sodium dodecyl sulfate, 0.05 % bromophenol blue) and run at 300 V for 300 min. Small RNA between 19 and 26 bp was cut out and used for library preparation. RNA-seq: . Poly (A) capture libraries were generated at the Max Planck-Genome-Centre Cologne (Germany). According to the facility's procedure, stranded TruSeq RNA-seq libraries were prepared as described in (Sultan et al., 2012), using 3 μg of rRNA-depleted RNA as input. RNA sequencing was performed with an Illumina HighSeq2500 and 25 million single-end reads/sample and 150 bp read length at the Max Planck Genome Centre Cologne (Germany).   miRNA-seq: Small RNA sequencing libraries were generated using the Small RNA v1.5 Kit (Illumina), following the manufacturers protocol. miRNA sequencing was performed with an Illumina HighSeq2500, single-end reads and 100 bp read length at the Max Planck Genome Centre Cologne (Germany).
attributes.tag: GEO Accession
attributes.value: GSM3317087