GSM1438894
GSM1438894: 29; Homo sapiens; RNA-Seq
For each participant 20 mL of peripheral blood was collected into heparinized tubes. Enrichment for neutrophil was performed by a Dextran-Ficoll sedimentation and centrifugation method60. Briefly, 20 mL of peripheral blood was mixed with  Dextran-RPMI media in a 4:1 ratio. After 40 minutes incubation at RT, the upper layer (white blood cell-rich plasma) was layered onto Ficoll (aspirate: Ficoll of 2:1 ratio) and centrifuged for 15 min at 2500 rpm. To remove the remaining erythrocytes, the cell pellet was treated with 2 mL ddH2O for 10-15 seconds, after which 25 mL of RPMI was added. The suspension was centrifuged for 5 min at 1400 rpm and the cell pellet was resuspended in 20 mL phenol red-free RPMI-1640. An aliquot of this suspension was used to check presence of dead cells with trypan blue staining.  This preparation contained > 98% neutrophils. Total RNA was isolated with RNeasy Mini Kit (Qiagen) following manufacturer’s protocol. Two rounds of RNase-free DNase I digestion (Qiagen) was performed to eliminate genomic DNA during the extraction process. RNA concentrations were determined using NanoDrop 2000 (Thermo Scientific). The quality and integrity of the RNA was determined using the RNA 6000 Pico chip on 2100 Bioanalyzer (Agilent Technologies). The median RNA integrity number (RIN) for the 4 samples was 8.05. RNA libraries were constructed using 1 µg of total RNA with TruSeq stranded mRNA Sample Preparation kit (Illumina, USA) following the manufacturer’s protocol.
GEO
GSM1438894
GEO Accession
GSM1438894