RNA-Seq of Takifugu rubripes:testis
Total RNA was extracted from the sample by TRIzol method or CTAB method, and 18-30nt bands were selected by polyacrylamide gel (PAGE) electrophoresis, and small RNA was recovered. The 3' linker and 5' linker were ligated respectively, and then the small RNA ligated with the two linkers was subjected to reverse transcription and PCR amplification. Finally, a band of about 140bp was recovered and purified by using PAGE gel, and dissolved in EB solution to complete the library construction. The quality and yield of the constructed library were detected by Agilent2100 and ABI Step One Plus Real-time PCR system (Life Technologies), and sequenced on the computer.