Full-length transcriptome sequencing of Lysiosquillina maculata, Odontodactylus Japonicus, Odontodactylus scyllarus, Oratosquilla oratoria
The mantis shrimp were temporary reared for a week and then dissected to obtain muscle, gonad, compound eye, and viscus. And the mantis shrimp were placed on ice for cry-mo-anesthesia before dissection to minimize the suffering. Meanwhile, the obtained tissues were frozen with liquid nitrogen and stored at -80C for total RNA extraction of individual tissue. The total RNA extracted from each tissue is mixed equally into one total RNA, which was then tested for integrity, purity, and quantification. We remove rRNA from the total RNA that has passed the quality test to obtain purified mRNA and then clarified it no less than three times. And the above mentioned mRNA was reverse-transcribed into cDNA using SMARTer PCR cDNA Synthesis Kit and then amplified the cDNA using KAPA HiFi PCR Kit. The amplification products were purified and eluded, and the optimal size (approximately 0.5-6 kb) cDNA fragments were subjected to damaged repair, blunt-end ligation, and SMRTbell adaptor addition using SMRTbell Template Prep Kit to complete the construction of SMRT sequencing library. Subsequently, the library template and enzyme complexes were transferred to the PacBio RSII platform for real-time FL transcriptome sequencing of mantis shrimp.A