Transcription profiling of human peripheral blood mononuclear cells, or whole blood were isolated 0, 30 min, 6 h, 24 h, and 7 day after intravenous endotoxin challenge vs controls

ABSTRACT: Peripheral blood mononuclear cells were isolated 0, 30 min, 6 h, 24 h, and 7 day after intravenous endotoxin challenge using cell preparation tubes (Vacutainer® CPT). The PBMC layer was lysed with RLT buffer, homogenized, and then stored at –80°C. Total RNA was extracted as per RNeasy‚ Midi protocol (Qiagen). Double stranded cDNA was synthesized from total RNA (5 to 20 µg) using 7-d(T)24 primer and SuperScript‰ Double-Stranded cDNA Synthesis Kit and purified by phase lock gel-phenol/chloroform extraction followed by ethanol precipitation. Biotin-labeled cRNA was prepared by in vitro transcription using HighYield‰ RNA Transcript Labeling Kit followed by fragmentation with 5X fragmentation buffer. Fragmented cRNA (10 µg) was hybridized to Affymetrix Hu95Av2 oligonucleotide probe arrays for 1

ORGANISM(S): Homo sapiens

SUBMITTER: Anthony Suffredini 

PROVIDER: E-GEOD-3026 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress