GSM1909037: cec-4_2; Caenorhabditis elegans; RNA-Seq
Extraction of RNA was performed according to the WormBook protocol (Stiernagle, 2006). Total RNA was purified using RNeasy kit (QIAGEN 74104) including DNase treatment. Depletion of ribosomal RNA was done for 5 µg of total RNA with Ribo-Zero™ Margnetic Gold Kit (Epicentre MRGZG12324) and further concentrated with RNA Clean & Concentrator™ kit (Zymo Research R1015) according to corresponding manufacturer’s instructions. From the depleted RNA 50ng were used for library preparation with the ScriptSeq v2 RNA-seq Library preparation kit (Epicentre). Equimolar pools of 3 samples were created and loaded on an Illumina HiSeq v3 flowcell using a cBot. Sequencing was performed on a HiSeq 2500 sequencer for 51 cycles running RTA 1.17.21.3. Samples were demultiplexed and FastQ files were generated using blc2fastq-1.8.4.