rumen sample
total RNA was extracted from tissue samples. Nanodrop2000 was used to detect the concentration and purity of the extracted RNA, agarose gel electrophoresis was used to detect the RNA integrity, and Agilent2100 was used to determine the RIN value. mRNA is isolated from total RNA. After that, the mRNA was segmented, the cDNA was synthesized by reverse transcription, and the adaptor was connected to Illumina Hiseq for computer sequencing.1. Library enrichment, PCR amplification of 15 cycles;2. TBS380 (Picogreen) quantitative, according to the ratio of data mixing machine;3. Bridge PCR amplification was performed on the cBot to generate clusters;4. Illumina Hiseq sequencing platform for 2*150bp sequencing.