RNA-seq of Portanus restingalis
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 repaired at the end, added single nucleotide A (adenine) and connected with adapters. The suitable fragments are selected for the PCR amplification as templates. After the construction of the library, Qubit2.0 and Agilent 2100 are used to detect the concentration and insert size of the library, and Q-PCR method is used to quantify the effective concentration of the library to ensure the quality of the library.  At last, the library is sequenced using Illumina NovaSeq 6000.