title: The 454 GS-FLX sequencing data of Sitobion avenae Fabricius transcriptome
design: Total RNA was isolated using Trizol reagent (Invitrogen, USA) and RNA precipitation buffer according to the manufacturer’s instructions . It was treated with RNase-free DNase I for 30 min at 37C to remove residual DNA, and mRNA was isolated from DNA-free total RNA using a PolyA Tract Kit (Promega, USA) according to the manufacturer’s instructions. In total 500 ng of mRNA isolated from the total RNA, was fragmented by ZnCl2 solution, and then purified and condensed with RNeasy MinElute RNA Cleaning up Kit (Qiagen, Germany). The fragmented RNA was eluted from the membrane of the spin tube with 10 μl of RNase-free water, and reverse transcribed into first strand cDNA with a random primer and AMV reverse transcriptase. The second strand of cDNA was synthesized by DNA polymerase I, RNase H and ligase enzyme mixtures. After being blunted and appended with an Adenine base overhang at the 3' end, the double-stranded cDNA were ligated with GS-FLX sequencing adaptors. The fragments shorter than 500 bp were removed by Ampure Beads according to the manufacturer’s instructions.