the full lenth transcriptome of Hu sheep testis
To increase the number of testis-specific peptide identifications used for protein inference, a cDNA library was constructed from a pooled RNA sample of 15 testis tissues (M0, M3, M6L, M6S and M12, n = 3 for each group) and subjected to full-length transcriptome sequencing on the PacBio RS II platform by Genepioneer Biotechnologies Inc. (Nanjing, China). Briefly, total RNA was extracted from the frozen testis tissues with the RNAsimple Total RNA Extraction Kit (TIANGEN, Beijing, China, DP419) following the manufacturers instruction. The concentration and integrity of RNA were assessed using NanoDrop 2000 (Thermo Scientific, MA, USA) and agarose gel electrophoresis. RNA was reverse-transcribed into cDNA using SMARTer TMPCR cDNA Synthesis Kit (Clontech Inc., Mountain View, CA, USA). BluePippin was used to screen full-length cDNA fragments and three types of insert libraries (1-2 kb, 2-3 kb, and 3-6 kb) were constructed. Subsequently, the full-length cDNA was amplified, end-terminal repaired, connected with SMRT dumbbell adapters and digested by exonucleases. Finally, the sequencing libraries were obtained after a secondary screening using BluePippin.