title: GSM5272997: CGP37157 treated C.elegans rep1 [CGP_rep1_02_12]; Caenorhabditis elegans; RNA-Seq
library_construction_protocol: control and treated worms were collected with distilled H2O, transferred to a Falcon tube and washed three times to eliminate eggs or larvae left in the plates. Then they were transferred to an eppendorf tube, the rest of the supernatant was carefully removed and three freezing-unfreezing cycles in liquid nitrogen were performed.  Then, 10-20 silica beads and 500 µl of TRizolTM were added to each frozen sample, samples were homogenized using a beads homogenizer for 1 minute and incubated in ice for 5 more minutes. Then 100µl of chloroform were added to each sample, incubated for 3 minutes and centrifuged at 4ºC, 15 minutes at 15.000 rpm. The upper phase was then transferred to another eppendorf tube and finally, a solution 1:1 (v/v) of the sample and Ethanol 70% was prepared. For RNA purification, the QIAGEN RNEasy Mini Kit was used. 1 microgram RNA was prepared using TruSeq Stranded mRNA kit, following Illumina instructions. RNA quality and quantity was assesed using  a Fragment Analyzer (AATI) and Qubit (ThermoFisher). Libraries run on a HiSeq1500 Illumina in 100 cicles paired-end reads.