mRNA seq for the study of spermatogenesis and fertilization
Total RNA was purified from target tissues conventionally and evaluated by Agilent 2100 Bioanalyzer (Agilent Technology, Santa Clara CA 95051). From this qualified RNA sample, a normalized cDNA library (insert size: 300-400 bp) was constructed by a process with TrueSeq RNA Sample Prep Kit (Illumina, San Diego, CA 92122) followed by Duplex Specific Nuclease normalization (Illumina) according to the manufacturer’s instruction. Paired-end sequencing (100 bp read x 2) was carried out by Illumina HiSeq2000 and a set of raw read data was produced after base calling and Chastity filtering (CASAVA ver.1.8.1., Illumina).