GSM9182205: CCR7- memory CD8+ T cells, IL-15 + Ionomycin, 12hours, rep1; Homo sapiens; RNA-Seq
Total RNA was extracted using TRIZOL reagent. An RNA library was prepared using a QuantSeq 3' mRNA-seq Library Prep Kit (Lexogen). An individual set of 500 ng total RNA was prepared, an oligo-dT primer containing an Illumina-compatible sequence at its 5' end was hybridized to the RNA, and reverse transcription was performed. After degradation of the RNA template, second strand synthesis was processed using a random primer with an Illumina-compatible linker sequence at its 5' end. Magnetic beads were used to purify the double-stranded library, and to eliminate reactive components. The library was amplified for the addition of complete adapter sequences needed for cluster generation and, finally, the amplified library was purified from PCR components.