title: Bulkseq_SCI2
design: Total RNA was extracted from tissue using TRIzol Reagent and assessed for quality. Then RNA quality was determined by 5300 Bioanalyser (Agilent) and quantified using the ND-2000 (NanoDrop Technologies).High-quality RNA samples (OD260/280 = 1.82.2, OD260/230  2.0, RIN  6.5, 28S:18S  1.0, > 1g) were used for library construction. RNA purification, reverse transcription, library preparation, and sequencing were performed by Shanghai Majorbio Bio-pharm Biotechnology Co., Ltd. following Illumina protocols. The RNA-seq library was prepared using 1g total RNA, including mRNA isolation with polyA selection, cDNA synthesis with random hexamer primers, and end-repair with 'A' base addition. Size-selected libraries (300 bp) were PCR amplified and sequenced on a NovaSeq 6000 sequencer (2  150bp)