Illumina NovaSeq 6000 paired sequencing of C2_ 1 (KAS24111175) (KAE24087921)
Eosinophils were differentiated from bone marrow cells in vitro (BMDE), and those were cultured with or without 10 mM acetate (C2) for 4 hours. For RNA extraction, BMDEs were lysed with Qiazol, and chloroform was added in a 5:1 ratio. Following centrifugation, the clear upper phase was collected, and RNA was precipitated by adding an equal volume of 100% isopropanol along with 20 ug of glycogen. After another round of centrifugation, the resulting pellet was washed with 75% ethanol and allowed to air-dry. Lastly, RNA was dissolved in nuclease-free water. For library construction, ribosomal RNAs were removed using NEBNext rRNA Depletion Kit v2, and the cDNA library was synthesized using NEBNext Ultra II Directional RNA Library Prep kit according to the manufacturer&#39;s protocols.