PMA-3 (CRX2082764)
Total RNA was isolated from the specified biological materials using TRIzol Reagent (Thermo Scientific, cat. #15596026) following the manufacturer's recommended protocol. Subsequent to RNA extraction, genomic DNA was removed by treating the samples with DNase I (NEB, cat. #M0303L) to eliminate potential DNA contamination. The purity of the extracted RNA was assessed by measuring the A260/A280 ratio using a Nanodrop TM OneC spectrophotometer (Thermo Scientific). The integrity of the RNA was verified using the LabChip GX Touch system (Revvity). The concentration of the qualified RNA was then determined using a Qubit 3.0 fluorometer with the QubitTM RNA Broad Range Assay kit (Thermo Scientific, cat. #Q10210). For library preparation, the total RNA was used as input for RNA sequencing library preparation utilizing the KCTM mRNA Library Prep Kit (Seqhealth Tech. Co., Ltd., Wuhan, China), according to the manufacturer's instructions. The library preparation included the enrichment of PCR products corresponding to fragments ranging from 200 to 500 base pairs. The enriched libraries were quantified and sequenced on a DNBSEQ-T7 platform (MGI) using the PE150 sequencing mode to generate paired-end reads.