GSM4175020: A3 blood NKG2a/cHighCD16-; Chlorocebus sabaeus; RNA-Seq
Cells were directly collected in a lysis buffer (RLT plus) containing TCEP. RNA was isolated using the RNeasy Plus® Micro Kit (74034, Qiagen). RNA integrity was verified with the Agilent Bioanalyzer. RNA from the different NK subsets were treated for library preparation using the Truseq Stranded mRNA sample preparation kit (Illumina, San Diego, California) according to manufacturer's instructions. Directional libraries were prepared using the Smarter Stranded Total RNA-Seq kit- Pico Input Mammalian following the manufacturer's instructions (Clontech 635005). The quality of all libraries was checked with the DNA-1000 kit (Agilent) on a 2100 Bioanalyzer and quantification was performed with Quant-It assays on a Qubit 3.0 fluorometer (Invitrogen).  Clusters were generated for the resulting libraries, with Illumina HiSeq SR Cluster Kit v4 reagents. Sequencing was performed with the Illumina HiSeq 2500 system and HiSeq SBS kit v4 reagents. Runs were carried out over 100 cycles, including seven indexing cycles, to obtain 100-bp single-end reads. Sequencing data were processed with Illumina Pipeline software (Casava version 1.9).
GEO
GSM4175020
GEO Accession
GSM4175020