title: GSM1697877: Starved_L1_daf16_A; Caenorhabditis elegans; RNA-Seq
library_construction_protocol: RNA was prepared using TRIzol (Invitrogen) according to the manufacturer's protocol with minor modifications. One milliliter of TRIzol was used per sample, and homogenization was supplemented with 100 μL of acid-washed sand. Poly-adenylated mRNA was isolated from total RNA using Dynal oligo(dT) magnetic beads (Invitrogen) according to the manufacturer's protocol. Two hundred nanograms of poly-adenylated RNA was used in a Tobacco Acid Pyrophosphatase reaction (Epicentre) according to the manufacturer's protocol in order to remove 5′ caps. The product was purified with phenol:chloroform extraction and ethanol precipitation using GlycoBlue as a coprecipitant (Ambion). This purified product was then used in the RNase III fragmentation reaction at the beginning of the Solid Total RNA-Seq Kit Whole Transcriptome protocol (Applied Biosystems). The manufacturer's instructions were followed for the remainder of the library preparation process. Fragmentation efficiency was analyzed with the BioAnalyzer (Agilent), and one-half of each sample (corresponding to ∼100 ng of RNA) was used for adapter ligation. cDNA was gel-purified to capture inserts of RNA fragment size 100–200 nt. Twelve PCR cycles were used to amplify the libraries.
identifiers.namespace: GEO
identifiers.id: GSM1697877