GSM2440681: WD in ecdysone_9h replicate 3; Drosophila melanogaster; RNA-Seq
RNA was isolated using a Qiagen RNeasy Mini kit. Samples were lysed by vortexing for 20-30sec in Buffer RLT + 2-mercaptoethanol on ice. Manufacturer's instructions were followed. Ethanol precipitation was subsequantly performed to achieve greater purification. mRNA was isolated from 1ug of total RNA using poly-dT enrichment with the NEBNext Poly(A) mRNA Magnetic Isolation Module. Final elution was in 15ul of 2x first strand cDNA synthesis buffer. NEBNext Ultra Directional RNA Library Prep Kit was used after chemical fragmentation of the sample at 94C for 15min. Custom adaptors were used for ligation and then depleted using an XP bead purification (Beckman Coulter) with a ratio of 1:1. Indexing was achieved during PCR enrichment using amplification primers carrying the index sequence. Two more XP bead purifications were done, and then the samples were quantified with Qubit dsDNA HS Assay Kit (Invitrogen).
GEO Accession
GSM2440681