GSM4459773: 26_349-09_Day_0; Macaca mulatta; RNA-Seq
NK cells were sorted directly into cold (4ºC) RPMI. The sorted NK cells were pelleted (500 x g, for 10 minutes) and lysed by vortexing for 1 minute in cold supplemented RLT buffer (RLT + β-MeOH) using the following ratio: 50µL cells in R10 and 350µL RLT buffer, 4ºC. Samples were then immediately frozen at -80ºC. RNA was extracted from these samples using the RNeasy Micro kit (Qiagen) with on-column DNase digestion. RNA quality was assessed using an Agilent Bioanalyzer. Five (5) nanograms of total RNA was used as input for cDNA synthesis using the Clontech SMART-Seq v4 Ultra Low Input RNA kit (Takara Bio) according to the manufacturer's instructions. Amplified cDNA was fragmented and appended with dual-indexed bar codes using the NexteraXT DNA Library Preparation kit (Illumina). Libraries were validated by capillary electrophoresis on an Agilent 4200 TapeStation, pooled at equimolar concentrations, and sequenced on an Illumina HiSeq3000 at 100SR, yielding 20-25 million reads per sample.
GEO Accession
GSM4459773