GSM5461929: PMN - donor_28 timepoint_1; Homo sapiens; RNA-Seq
Polymorphonuclear cells were isolated from peripheral blood after gradient centrifugation using Lymphoprep (1.077 g/ml, Alere Technologies), carefully harvesting the top part of the bottom layer. Remaining red blood cells were lysed with RBC lysis buffer (BD Bioscience) and clean PMNs were washed twice with PBS. PMNs were directly lysed in TRI-reagent (Sigma-Aldrich). Total RNA was isolated using the RNeasy mini kit (QIAGEN). An Illumina sequencing library was prepared using the NEBNext Ultra II Directional RNA Library Prep Kit fo Illumina. 250ng of total RNA were used for library preparation.