title: RNA-seq of Usuironus limbifer
design: After the total RNA extraction and DNase I treatment, magnetic beads with Oligo (dT ) are used to isolate mRNA. Mixed with the fragmentation buffer, the mRNA is fragmented into short fragments. Then cDNA is synthesized using the mRNA fragments as templates. Short fragments are purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments are connected with adapters. The suitable fragments are selected for the PCR amplification as templates. During the QC steps, Agilent 2100 Bioanaylzer and ABIStepOnePlus Real-Time PCR System are used in quantification and qualification of the sample library. At last, the library is sequenced using Illumina.