title: GSM9162326: Early stage wild-type corneal endothelium, replicate 2; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted from tissues or cells using TRIzol reagent (Invitrogen, USA) according to the manufacturer's instructions. RNA purity and concentration were assessed using a NanoDrop 2000 spectrophotometer (Thermo Scientific, USA). RNA integrity was evaluated with an Agilent 2100 Bioanalyzer (Agilent Technologies, USA) to ensure sample quality met the requirements for library construction. RNA-seq libraries were constructed using the NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina (Cat. No. NEB#E7330S, NEB, USA) following the manufacturer's instructions. The library preparation workflow included mRNA enrichment or rRNA depletion, RNA fragmentation, first- and second-strand cDNA synthesis with strand specificity, end repair, adapter ligation, and PCR amplification. Library quality and fragment size distribution were assessed using a Bioanalyzer to ensure suitability for sequencing.