GSM9081722: bGV_oocytes,SAC-Seq treated (A site modified) sampes_Replicate2; Bos taurus; RNA-Seq
samples were lysed in TRIsolution (Sigma), followed by phase separation with chloroform. The aqueous phase was collected, and RNA was precipitated with isopropanol, washed with ethanol, and resuspended in nuclease-free water. Total RNA was directly subjected to Selectively Allyl-Chemically Labeled m6A Sequencing to achieve single-nucleotide resolution detection of m6A modifications. The RNA underwent selective allyl-chemical labeling of m6A sites, followed by reverse transcription using a modified primer designed to introduce mutation signatures indicative of m6A presence. cDNA libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit (New England Biolabs)