alias: GSM2465956
title: GSM2465956: Five hours post fertilization replicate 2013_25; Mnemiopsis leidyi; RNA-Seq
library_construction_protocol: Total RNA was harvested for 100-200 embryos per stage with the Trizol reagent. Dynabeads were used to extract mRNA. Libraries were prepared using ScriptSeq V2 (Epicenter) with some modifications. Briefly, instead of the manufacturer’s recommended 5 minute fragmentation, we performed a 90 second fragmentation at 85oC, followed by 15 rounds of PCR amplification. The quality of the resulting libraries was tested using High Sensitivity DNA Chip Kits (Agilent). After quality testing, the libraries were subjected to targeted size selection using Pippin Prep (Sage Science) to a range encompassing 350bp to 600bp. After size selection, the samples were cleaned up with PCR minElute columns from Qiagen to both concentrate the DNA and to remove the EtBr from the Pippin Prep. The samples were then run again on High Sensitivity DNA Chips to confirm quality. The resulting high quality libraries were pooled and sequenced with an Illumina High Seq sequencer (Illumina).
identifiers.namespace: GEO
identifiers.id: GSM2465956