title: GSM2460232: fog1_XX_youngadult_rep2; Caenorhabditis elegans; RNA-Seq
library_construction_protocol: Total RNA isolation protocol: Ten volumes of Trizol was added to worm collections. Total RNA was extracted by freeze cracking the samples (briefly, samples were flash frozen in liquid nitrogen and then thawed by placing in a 37C water bath) 3-5 times. Total RNA was isolated according to the manufacturer's (Invitrogen) Trizol protocol. Purified Total RNA was cleaned up on a Qiagen RNeasy column. cDNA Synthesis Protocol: mRNA was isolated from 500ng-10ug of total RNA using Sera-Mag Oligo(dT) beads (ThermoScientific). As in staging protocol: worms were synchronized by bleaching gravid adults from male-heavy plates. At least 750 young adult worms were hand picked for each replicate.  RNA isolation protocol: RNA was extracted by freeze cracking embryos by freezing in liquid nitrogen and thawing at 37C water bath five times. RNA was isolated according to the manufacturer's (Invitrogen) Trizol extraction protocol. Purified total RNA was cleaned up through Qiagen RNeasy column.  cDNA synthesis protocol: mRNA was purified from 5-10 ug of total RNA using Sera-Mag Oligo(dT) beads (ThermoScientific). cDNA preparation was done in the presence of dUTP to prepare stranded RNA-seq libraries as in PMID:19620212, cDNA synthesis was performed in the presence of dUTP in order to prepare stranded mRNA-seq libraries. cDNA was ligated to Illumina or home-made multiplexed adapters and amplified by PCR. Library DNA between 250-500 bp in size was gel purified.
attributes.tag: GEO Accession
attributes.value: GSM2460232