Genome-scale transcript analyses in human neutrophils.
Transcriptome analyses of single- and multicellular organisms have changed fundamental understanding of biological and pathological processes across multiple scientific disciplines. Over the past 5 yr, studies of polymorphonuclear leukocyte (or neutrophil) (PMN) gene expression on a global scale have provided new insight into the molecular processes that promote resolution of infections in humans. Herein, we present methods to analyze gene expression in human neutrophils using Affymetrix oligonucleotide microarrays, which include isolation of high-quality RNA, generation and labeling of cRNA, and GeneChip hybridization and scanning. Notably, the procedures utilize commercially available reagents and materials and thus represent a standardized approach for evaluating PMN transcript levels.
Kobayashi
Scott D
SD
Department of Microbiology, Molecular Biology, and Biochemistry, University of Idaho, Moscow, ID, USA.
Sturdevant
Dan E
DE
DeLeo
Frank R
FR
D001781:Blood Component Removal / Q000379:methods; D002469:Cell Separation; D020869:Gene Expression Profiling / Q000379:methods*; D015894:Genome, Human; D006801:Humans; D007091:Image Processing, Computer-Assisted; D009504:Neutrophils / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011786:Quality Control; D018075:RNA, Complementary / Q000138:chemical synthesis / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism
D018075:RNA, Complementary; D012333:RNA, Messenger