GSM3263613: Lim3_knockdown; Drosophila melanogaster; RNA-Seq
Total RNA was extracted from embryos of 0- to 16-hour (100–200 mkg)  using ExtractRNA (Evrogen). The poly-A containing mRNA fraction was isolated from total RNA samples using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB) according to the manufacturer's protocol. The sequencing libraries were constructed from the poly-A containing mRNA fraction using NEB Next Ultra Directional RNA Library Prep Kit for Illumina (NEB) following the manufacturer's protocol with some modifications, namely fragmentation time was 94°C for 15 m; Agencourt Ampure XP beads (Beckman Coulter, Inc.) were used to purify ds cDNA >200bp after adapter ligation.  The obtained sequencing libraries of fragment size between 250 and 400bp were purified by gel extraction using Qiagen Gel Extraction (Qiagen).