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Genome-Scale Transcript Analyses with Human Neutrophils

  • Scott D. Kobayashi
  • Daniel E. Sturdevant
  • Frank R. DeLeo
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1124)

Abstract

Transcriptome analyses of single- and multicellular organisms have changed fundamental understanding of biological and pathological processes across multiple scientific disciplines. Over the past decade, studies of polymorphonuclear leukocyte (PMN or neutrophil) 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.

Keywords

Neutrophil Microarray Gene expression Affymetrix Transcript Phagocyte Polymorphonuclear leukocytes 

Notes

Acknowledgments

This work was supported by the Intramural Research Program of the National Institutes of Allergy and Infectious Diseases, NIH.

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Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • Scott D. Kobayashi
    • 1
  • Daniel E. Sturdevant
    • 2
  • Frank R. DeLeo
    • 3
  1. 1.Department of Microbiology, Molecular Biology, and BiochemistryUniversity of IdahoMoscowUSA
  2. 2.Genomics Unit, Rocky Mountain Laboratories, National Institute of Allergy and Infectious DiseasesNational Institutes of HealthHamiltonUSA
  3. 3.Laboratory of Human Bacterial Pathogenesis, Rocky Mountain LaboratoriesNational Institute of Allergy and Infectious Diseases, National Institutes of HealthHamiltonUSA

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