RNA-seq of Xestospongia muta
1) Isolate the Total RNA from sample.2) Eliminate DNA contamination using DNase.3) Use ribo-zero RNA removal kit to remove ribosome RNA and purify total coding and non-coding RNA.4) Randomly fragment purified RNA for short read sequencing.5) Reverse transcribe fragmented RNA into cDNA.6) Ligate adapters onto both ends of the cDNA fragments.7) After amplifying fragments using PCR, select fragments with insert sizes between 200-400 bp. For paired-end sequencing, both ends of the cDNA is sequenced by the read length.