RNA-Seq of testis: microplastic exposed
Total RNA was isolated using the TRIzol reagent, followed by purification with PureLink RNA Mini Kit columns according to the manufacturers protocol. RNA concentration and purity were assessed using a Qubit Fluorometer, and integrity was confirmed on an Agilent TapeStation, with all samples showing an RNA Integrity Number (RIN) greater than 9.5. High-quality RNA samples from control and treatment groups (n = 2 per group) were used for library preparation with the KAPA RNA HyperPrep Kit. RNA sequencing (RNA seq), a high-throughput technique for transcriptome profiling, was performed on the Illumina NovaSeq 6000 platform, enabling accurate quantification of gene expression, identification of novel transcripts, and analysis of alternative splicing events.