GSM4610491: NK_anti_CD28H_6; Homo sapiens; RNA-Seq
Total RNA were extracted using  the RNeasy Mini kit (Qiagen) following manufacturer's protocol. RNA quality was assessed using an Agilent TapeStation 4200 (Agilent Technologies, Santa Clara, CA, USA). The library preparation was done using TruSeq Stranded mRNA Sample preparation kit (Illumina inc) with 250ng of total RNA. Enriched mRNA using oligodT beads were fragmented using enzymatic fragmentation and used for cDNA synthesis before double stranded cDNA were purified (AMPure XP, Beckman Coulter). The cDNA was end-repaired, 3' adenylated and Illumina sequencing adaptors ligated onto the fragments ends, and the library was purified (AMPure XP). The mRNA stranded libraries were pre-amplified with PCR and purified (AMPure XP). The libraries size distribution was validated and quality inspected on a BioAnalyzer 4200 tapeStation (Agilent Technologies). Single end sequencing was carried out on a NextSeq500 instrument (75 cycles) according to the manufacturer instructions (Illumina Inc.).
GEO Accession
GSM4610491