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Isolation and Characterization of MicroRNAs of Human Mature Erythrocytes

  • Carolyn Sangokoya
  • Gregory LaMonte
  • Jen-Tsan Chi
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 667)

Abstract

Human mature erythrocytes are terminally differentiated cells that have lost their nuclei and organelles during development. Even though mature erythrocytes lack ribosomal and other large-sized RNAs, they still retain small-sized RNAs. We have recently shown that there are abundant and diverse species of microRNAs in mature erythrocytes through the use of several different techniques, including northern blot, miRNA microarray, and real-time PCR. Furthermore, fractionation and genomic analysis has revealed that erythrocyte microRNA expression is different from that of reticulocytes or leukocytes and that mature erythrocytes contribute the majority of microRNA expression in whole blood. Therefore, global analysis of microRNA expression in circulating erythrocytes has the potential to provide mechanistic insights into erythrocyte biology and erythrocyte-related disorders. Here, we have provided the detailed methods for isolating and characterizing the microRNAs from human mature erythrocytes to enable such researches into human diseases involving erythrocytes.

Keywords

Methylene Blue Wash Buffer Acridine Orange Polycythemia Vera Erythroid Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We thank the Duke microarray facility and members of the Chi lab for technical assistance and constructive feedback and the Telen lab for assistance with sample collection. This research was funded by NIH R21DK080994 and Roche Foundation for Anemia Research (RoFAR).

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Carolyn Sangokoya
    • 1
  • Gregory LaMonte
    • 1
  • Jen-Tsan Chi
    • 1
  1. 1.Department of Molecular Genetics and Microbiology, The Institute for Genome Sciences and PolicyDuke University School of MedicineDurhamUSA

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