GSM4566997: Unstimulated Malian Adults [R9]; Homo sapiens; RNA-Seq
Cells were isolated and kept in RNAprotect buffer (Qiagen) at -80 degree before RNA was isolated from the samples. RNA was isolated using RNAeasy kit according to manufacturer's instruction. Isolated RNA's were checked for quality and quantified using Agilient Bioanalyzer. RNA's with RIN values greater than 7 were used for all the analysis. cDNA was prepared from 10 ng total RNA using the Ovation® RNA Seq System V2 (Tecan) according to manufacturer's instructions.  This method employs both poly-T and random primers so that both poly-adenylated and non-poly-adenylation messages are included.  The cDNA product was end-repaired using the NEBNext End Repair Module (New England Biolabs). RNA-Seq libraries were prepared using 1 g of end-repaired cDNA using the TruSeq Stranded RNA Kit (Illumina), however due to the method of amplification the libraries are not stranded.  Unique dual-indexed barcode adapters were applied to each library. Libraries were pooled in an equimolar ratio for sequencing.  The pooled libraries were sequenced on one lane of an S4 flow cell on a NovaSeq 6000 using version 1 chemistry to achieve a minimum of 49 million 150 base read pairs.  The data was processed using RTA version 3.4.4 and BWA-0.7.12.
GEO Accession
GSM4566997