GSM9326179: Head and Neck case 11; Homo sapiens; RNA-Seq
Total RNA was isolated from human tissue specimens using the AllPrep DNA/RNA Mini Kit (Qiagen, Cat. No. 80204) in accordance with the manufacturer's protocol. Briefly, fresh or snap-frozen tissue was transferred to 2 mL tubes containing ceramic beads (Omni International, Cat. No. 19-646-3) and homogenized in RLT buffer using the Bead Ruptor 12 (Omni International) at 3-4 M/s for 5 seconds, repeated for one to two cycles to ensure complete disruption. Following a centrifugation, the resulting supernatant was processed through the kit's spin columns to sequentially purify genomic DNA and total RNA. RNA was eluted in RNase-free water, and concentration and purity were measured using a NanoDrop 8000 spectrophotometer (Thermo Fisher Scientific). RNA integrity was assessed using the Agilent TapeStation. Purified RNA samples were stored at -80°C until downstream analyses RNA libraries for RNA sequencing were prepared using Novogene and following the Illumina manufacturer's protocol. Paired-end sequencing was performed on an Illumina HiSeq 2500 platform following library quality control and poly(A) enrichment, using a read length of 150 base pairs.