Blood samples were obtained from healthy adults via venipuncture into vacutainer tubes. Blood was washed three times with PBS, centrifuged, and plasma and buffy coat removed. Leukocytes were then depleted from the sample using a Purecell® Leukocyte Reduction Filtration System. Reticulocytes (immature red blood cells) were removed in the CD71 positive fraction using CD71 microbeads with the autoMACS® Separator (Miltenyi Biotec). The erythrocyte purification protocol reflects methods previously described (Sangokoya et. al, Methods in Molecular Biology, 2010). RNA was extracted using the miRVana miRNA Isolation kit (Life Technologies). Total RNA-seq libraries were prepared with 1ug purified total RNA input using the TruSeq Stranded Total RNA Sample Prep Kit with Ribo‐Zero Globin (Illumina). Samples were muliplexed, libraries pooled, and run on two flowcells with the Illumina HiSeq 2000 sequencing system. Barcode IDs are as follows: RBC_1-CGATGT, RBC_2-TGACCA, RBC_3-ACAGTG, PBMC_1-GCCAAT, PBMC_2-CAGATC, PBMC_3-CTTGTA, D8_1-AGTCAA, D8_3-GTCCGC.
GEO Accession
GSM1830784