GSM1319921: XX females; Tachysurus fulvidraco; RNA-Seq
Gonad tissues were taken from each individual of XX females, XY males, YY super-males. Total RNA was isolated from each sample by a Qiagen miRNeasy Mini Kit (Qiagen, USA) according to the manufacturer’s protocol and standardized to a concentration of 1 μg/μL. RNA from the same type tissues was combined into one RNA pool, whose quality and quantity was measured with NanoDrop 2000 and Agilent 2100 Bioanalyzer. Three small RNA libraries were constructed with these three RNA pools using the TruSeq Small RNA Sample Preparation Kits. Briefly, total RNA was ligated with a 3' RNA adaptor and then with a 5' RNA adaptor. PCR amplification after reverse transcription was performed to enrich the fragments that had adaptors on both ends. Subsequently, the cDNA constructs were purified and enriched with 6% denaturing polyacrylamide gel electrophoresis to isolate the expected size fractions and eliminate dimerized adaptors. Finally, these three RNA libraries were sequenced using an Illumina/Solexa Genome Analyzer