antler growth center at 30d
Total RNA was extracted using TRIzol reagent (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer's protocol. Extracted RNA was treated with DNase I (Promega, Madison, WI, USA) to remove contamination. To ensure that RNA samples met sequencing requirements, their concentration and integrity were evaluated using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, USA), RNA Nano 6000 analysis kit and Bioanalyzer 2100 system (Agilent Technologies, Santa Clara, CA, USA), respectively. If 1.8 < OD260/OD280 < 2.0, RIN > 7.0 and r RNA 28S/18S  0.7, the extracted total RNA meets the quality requirements and can be used for the construction of small RNA libraries. Sequencing libraries were generated by using the NEBNext-Ultrafi RNA Library Preparation Kit for Illuminafi (New England Biolabs, Ipswich, MA, USA). Library quality was evaluated using a Bioanalyzer 2100 (Agilent Technologies). The miRNA libraries were sequenced using the HiSeqfi Rapid SBS kit v2 and the HiSeq 2500 platform