small RNA-Seq of mouse: adult sperm
The total RNA was extracted from sperm by using the Trizol method. The quality and quantity of RNA were determined by Nanodrop-1000. 400 ng total RNAs are used for small RNA library construction by using NEBNext Multiplex Small RNA Library Prep Set for Illumina (NEB, E7300, USA). The PCR amplification products were purified by DNA Clean & Concentrator-5 Kit (Zymo Research, D4003) followed by size selection using PinPin Prep (Sage Science, CDP 3010). The quality of the library is checked by Bioanalyzer 2100 (Agilent Technologies, Inc.). The high-throughput sequencing is completed by Illumina HiSeq 2500 sequencer and 12 million reads of raw data are obtained.