RNA-Seq of African ostrich chick:Spleen lymphocyte
Construction and sequencing of RNA library was provided by Annuo Youda Gene Co., Ltd (Zhejiang,China). After passing the quality inspectiontotal mRNA was isolated using Oligo-dT beads and broken into short fragments. A six-base random primer was used to synthesize the first-strand of the cDNA, and the second -strand of the cDNA was synthesized by adding buffers, dNTPs, RNaseH and DNA Polymerase I. Short cDNA fragments were purified by QIAQuick PCR Kit and eluted with EB buffer. The purified double-stranded cDNA was subjected to end-repairing, base A addition, and sequencing ligation, and then the target size fragment was recovered by agarose gel electrophoresis and PCR amplification was performed to complete the entire library preparation work. After the library was constructed, the effective concentration of the library was accurately quantified (the effective concentration of the library was greater than 10 nM) to ensure the quality of the library, and the qualified library was sequenced using the Illumina platform. The sequencing strategy was PE150.