title: Transcriptome analysis of the GIFT Strain of Nile Tilapia (Oreochromis niloticus) spleen in response to Streptococcus agalactiae ZQ0910
design: The Illunima HiSeqTM2000 technology was applied to determine the transcriptome sequence of Oreochromis niloticus spleen tissue and a comparative analysis of transcriptome date between the control and the inactivity vaccine of Streptococcus agalactiae ZQ0910 infected group was performed in this study.     The cDNA library was constructed according to the manufacturer’s instructions (Illumina/Hiseq-2000 RNA-seq). The mRNA molecules containing poly (A) were purified using Sera-mag Magnetic Oligo (dT) Beads from the RNA samples.A fragmentation buffer was added to break the mRNA into short fragments. Using these short fragments as templates, the first strand of cDNA was synthesized.The second strand of cDNA was synthesized using the buffers containing dNTPs, RNaseH, and DNA polymerase I. The synthesized cDNA was purified using the QiaQuick PCR extraction kit (Qiagen). The purified cDNA was connected with the sequencing adapters. A range of cDNA fragments (200*25 bp) were excised from 2% TAE-agarose gel (Certified Low-Range Ultra Agarose, Biorad) using QIAquick Gel Extraction Kit(Qiagen). The library was sequenced using the Illumina/Hiseq-2000 RNA-seq.