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Serial Analysis of Gene Expression Adapted for Downsized Extracts (SAGE/SADE) Analysis in Reticulocytes

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DNA and RNA Profiling in Human Blood

Part of the book series: METHODS IN MOLECULAR BIOLOGY™ ((MIMB,volume 496))

Abstract

Reticulocytes are the last stage of erythropoiesis before red blood cells (RBC). Although most of the RBC proteins have been characterized, little is known about expression profile of their mRNA during differentiation. Our goal was to initiate a genomic global solution to provide a transcriptional data base on which it will be possible to follow the erythroid differentiation and study RBC disorders. We used a modified protocol of serial analysis of gene expression (SAGE), the serial analysis of downsized extracts (SADE) which allows identification of genes from small amounts of mRNA and is thus an appropriate method starting from reticulocyte. SAGE does not depend on previous knowledge of genes. The method produces a set of cellular transcribed gene signatures (tags) and provides a comprehensive view of cells phenotypes by computational analyses.

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References

  1. Velculescu, V. E., Zhang, L., Vogelstein, B., et al. (1995) Serial analysis of gene expression. Science 270, 484–487.

    Article  CAS  PubMed  Google Scholar 

  2. Virlon, B., Cheval, L., Buhler, J. M., et al. (1999) Serial microanalysis of renal transcriptomes. Proc Natl Acad Sci USA 96, 15286–15291.

    Article  CAS  PubMed  Google Scholar 

  3. Bonafoux, B., Lejeune, M., Piquemal, D., et al. (2004) Analysis of remnant reticulocyte mRNA reveals new genes and antisense transcripts expressed in the human erythroid lineage. Haematologica 89, 1434–1438.

    CAS  PubMed  Google Scholar 

  4. Piquemal, D., Commes, T., Manchon, L., et al. (2002) Transcriptome analysis of monocytic leukemia cell differentiation. Genomics 80,361–371.

    Google Scholar 

  5. Quere, R., Manchon, L., Lejeune, M., et al. (2004) Mining SAGE data allows large-scale, sensitive screening of antisense transcript expression. Nucleic Acids Res 32,e163.

    Article  PubMed  Google Scholar 

  6. Beutler, E., Gelbart, T. (1986) The mechanism of removal of leukocytes by cellulose columns. Blood Cells 12, 57–64.

    CAS  PubMed  Google Scholar 

  7. Chomczynski, P., Sacchi, N. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162, 156–159.

    Article  CAS  PubMed  Google Scholar 

  8. Quéré, R., Manchon, L., Pierrat, F., et al. (2007) Rapid and accurate pyrosequencing of serial analysis of gene expression ditags. Application Note Nˆ4 Roche diagnostics.

    Google Scholar 

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Acknowledgments

We acknowledge the support of the Languedoc-Roussillon Génopole\({\circledR}\) for this work.

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Bonafoux, B., Commes, T. (2009). Serial Analysis of Gene Expression Adapted for Downsized Extracts (SAGE/SADE) Analysis in Reticulocytes. In: Bugert, P. (eds) DNA and RNA Profiling in Human Blood. METHODS IN MOLECULAR BIOLOGY™, vol 496. Humana Press. https://doi.org/10.1007/978-1-59745-553-4_20

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  • DOI: https://doi.org/10.1007/978-1-59745-553-4_20

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-934115-93-0

  • Online ISBN: 978-1-59745-553-4

  • eBook Packages: Springer Protocols

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