GSM8712045: P3-2; Homo sapiens; RNA-Seq
The frozen samples stored were thawed, and immediate steps were taken for RNA extraction. To ensure consistency, all samples were processed simultaneously to avoid any potential batch effects. RNA was extracted and purified from the patient specimens using the Direct-zol RNA Microprep Kit (Cat. #R2063), following the manufacturer's protocol. Nuclease-free commercially available water was used during the RNA isolation process. The quality and quantity of the extracted RNA were immediately assessed using Nanodrop UV measurements. Following the measurements, all isolated RNA samples were stored at -80° C until sequencing. An RNA-seq library was generated utilizing the Illumina mRNA sample prep kit (Cat. #RS-100-0801) according to the manufacturer's protocol in 3 technical replicates for each sample. The library constructs' size and concentration were verified with a bioanalyzer before sequencing on the Illumina NovaSeq6000. The raw reads' quality was evaluated to confirm the success of the library preparation and sequencing processes.