GSM8221222: NC1_Ribo; Sus scrofa; OTHER
Ribo-seq:In a nutshell, ribosome protected mRNA is obtained through MNase digestion and RNA purification. Briefly, the longissimus dorsi muscle samples were ground in liquid nitrogen and extracted with 800 μL of polysome lysis buffer (20 mM Tris pH 7.5, 150 mM NaCl, 5 mM MgCl2, 24 U/mL TurboDNase, 100 μg/mL cycloheximide, 1 mM dithiothreitol (DTT), 1% Triton X-100, and protease inhibitor mixture) for 30 min. Ribo-seq libraries were prepared according to reference with a minor modification. In total, 5 μg of ribosome-protected mRNA was used to prepare a library with the NEBNext Multiple Small RNA Library Prep Set for Illumina HiSeqX Ten. Briefly, adapters were added to both ends of Ribosome footprints (RFPs), followed by reverse transcription and PCR amplification.