Transcriptome analysis of the Spodoptera exigua challenged with active or inactivated Autographa californica
The mRNA was purified from the total RNA extracted from 5th instar larvae of S. exigua using     the Oligotex mRNA Midi Kit (QIAGEN, Germany). Then, double-stranded cDNA was synthesized using the SMART cDNA Library     Construction Kit (Clontech, USA). PCR products of cDNA were further purified using the QIAquick PCR Purification Kit (QIAGEN,     Germany) and checked for quality using the Agilent 2100 Bioanalyzer. Approximately 5 ug of cDNA was used for sequencing on a     Roche/454 GS-FLX Titanium instrument (Roche, Germany). A 1/8-plate sequencing run was performed at MACROGEN Co. (Korea)     following manufacturer's instructions. The raw 454 sequence files in SFF format were processed to remove low quality regions     and adaptor sequences using programs SeqClean (http://compbio.dfci.harvard.edu/tgi/software). The resulting sequences were     then screened against the NCBI UniVec database and E. coli genome sequences to remove possible contaminations of these     sequences. Sequences shorter than 100 bp were discarded. The processed 454 sequences were assembled into unigenes using the     iAssembler program (http://bioinfo.bti.cornell.edu/tool/iAssembler ).