GSM8247919: head and neck tumor before proton therapy human 4; Homo sapiens; RNA-Seq
Processing of the biomaterial was carried out under aseptic conditions using sterile tools. Total RNA was isolated from snap-frozen tissue after removal after homogenization with Teflon beads in QIAZOL (Qiagen), followed by purification and concentration with RNA Clean & Concentrator kit (Zymo Research), quantity was estimated using Qubit 2.0 device (Invitrogen, Life Technologies). After isolating total RNA from 1 μg, samples were prepared for sequencing on the Illumina platform. libraries using the TruSeq RNA kit Sample Prep Kit v2 with 10 cycle enrichment step in accordance with the manufacturer's recommendations. The quality of RNA and resulting libraries was checked using an Agilent 2100 Bioanalyzer capillary electrophoresis system (Agilent Tec. Inc., USA). The RIN parameter for RNA was at least 7. The concentration of the libraries was assessed using real-time polymerase chain reaction on the CFX96 Touch Real-Time PCR Detection System device (Bio-Rad, USA). The final libraries were pooled in equimolar proportions before sequencing on the Illumina HiSeq 2500 platform using 50-base paired-end reads.