title: GSM3145858: Ghrelin_TRT_Liver_male_6; Mus musculus; RNA-Seq
library_construction_protocol: Liver RNAs were extracted as previously described (Gachon et al, 2006) with classical methods using guanidium  thiocyanate lysis buffer. The RNA were quantified with Ribogreen (Life Technologies) and quality assessed on a Fragment Analyzer (Advances Analytical). Sequencing libraries were prepared from 250 ng total RNA using the TruSeq Stranded Total LT Sample Prep Kit (Illumina) with the Ribo-Zero Gold depletion set. The procedure was automated on a Sciclone NGS Workstation (Perkin Elmer). The manufacturer's protocol was strictly followed, except for the PCR amplification step. The latter was run for 12 cycles with the KAPA HiFi HotStart ReadyMix (Kapa BioSystems). This optimal PCR cycle number has been evaluated using the Cycler Correction Factor method as described previously (Atger et al., PNAS, 2015). Purified libraries were quantified with Picogreen (Life Technologies) and the size pattern was controlled with the DNA High Sensitivity Reagent kit on a LabChip GX (Perkin Elmer). Libraries were then pooled by 24 and each pool was clustered at a concentration of 7 pmol on 8 lanes of a v3 paired-end sequencing flow cell (Illumina). Sequencing was performed for 2 x 125 cycles on a HiSeq 2500 strictly following Illumina's recommendations.
attributes.tag: GEO Accession
attributes.value: GSM3145858